Early attempts to generate new cartilage tissue in vivo began in the 1970s, in Dr. W. T. Green’s lab at the Children’s Hospital [1]. While the initial work had not been successful, this led to the development of work by others including Drs. Burke and Yannas at Massachusetts General Hospital and M.I.T. to create tissue-engineered skin substitute supported by biocompatible materials. In 1998, the first bi-layered bioengineered skin substitute, Apligraf, was approved by the US Food and Drug Administration (FDA).

(About the feature image above: REGENERATIVE RELIQUARY an art piece bioprinted by Amy Karle 2016 )

The field of tissue engineering quickly evolved with pioneers like Dr. Robert Langer and Dr. Joseph Vacanti conceptualizing the idea of developing novel and customizable materials as scaffoldings for cell delivery [1]. Later, when Charles Hull introduced stereolithography, the evolution into bioprinting technology quickly followed suit and in 1999, scientists at the Wake Forest Institute of Regenerative Medicine (WFIRM) successfully printed the first synthetic scaffold of a human bladder, which led to the first lab-grown organs implanted into humans [2].

In the beginning, a major challenge faced in the bioprinting field was the lack of customizable biomaterials that could be easily printed for tissue engineering applications. Access to such technology was also prohibited by cost but all these problems quickly turned around in the last decade. Today, we can purchase a commercial desktop bioprinter for less than $10,000 USD or even build one ourselves for less than $1000. There is also an array of biomaterials that are photocrosslinkable, thermo-reversible, and readily printable at room temperature that would provide ideal printing and culture conditions to support different tissue engineering goals.

As research rapidly advances in the field of bioprinting and tissue engineering, we began to ask ourselves “What is the next bottleneck we must overcome to progress tissue-engineered product innovations into the clinic?”

It did not take too long for many tissue engineering and bioprinting researchers to realize that cells, often an essential component of the final product can potentially be a challenge. For autologous based products, this is highly dependent on the patient’s condition and therefore the quality and quantity of the cellular material will be highly variable patient-to-patient. However, for products that are being generated from allogeneic sources, such as young and healthy donors, there is the benefit of controlling critical quality attributes of the cellular input starting materials to ensure high quality and reproducibility of manufacturing process outcomes.

Peak MSC Demand

In Olsen et.al. ”Peak MSC” paper published in 2018 [3], the authors analyzed the growth in demand for mesenchymal stromal/stem cell (MSC) in many regenerative medicine products. Using bone tissue replacement as an example, Dr. Olsen projects a peak demand of 278 trillion (10¹²) cells per year based on 185,000 amputations performed annually in hospitals today. Just to provide some perspective, typical tissue engineering projects at research laboratories only utilize around 70 million cells and in vivo studies may require 100 million cells [3]. In order to generate billions or even trillions of cells for the production of tissue-engineered constructs, it would be necessary to have a “consistent and readily available supply of high quality, standardized and economical c-GMP cellular starting material to support this.” While tissue engineering applications has not moved as quickly as cell therapy into the clinic, various groups have steadily made progress on this front. According to the Alliance for Regenerative Medicine (ARM) 2019 annual report, there are 46 tissue engineering clinical trials (6 in Phase I, 23 in Phase II and 17 in Phase III) that are currently in the pipeline and the majority are targeted toward bone, cartilage, and skin tissue regeneration or replacement.

Projected near-term cell demands

In the nearer term, projected cell demands will increase for more mature applications like bone, cartilage, and skin tissue. The table below summarizes the current need-based on existing clinical trials and cell numbers used in animal models for bone and cartilage tissue.

In the case of cartilage tissue, typical defect size >3 cm2 are treated with cartilage tissue-engineered implants and the mean lesion size based on MACI’s (matrix-induced articular cartilage implantation) Phase 3 trial is around 4.8 cm2 [12], each patient will require ~14.4 million cells. To treat 10,000 patients every year, the annual cell demand (i.e. cell number required to treat patients) would be 144 billion cells which require a total manufacturing output of 308 billion cells. The manufacturing lot size requirements (i.e. how many cells you actually need to produce) are calculated based on cell recovery loss, viability drop, and cell harvest loss. If you are curious to learn how to calculate manufacturing lot sizes, please refer to this blog.

It would not be surprising that some of you might be thinking right now that the cell densities or numbers shown in Table 1 seem rather conservative. I would certainly agree but I also think this is dependent on the desired tissue type and application. In fact, many bioprinting and tissue engineering groups have reported the need for high cell densities in order to create functional tissues that mimic more of the in vivo environment. Researchers at Organovo reported cell densities of 150 million cells/mL of parenchymal or non-parenchymal cells to create a 3D liver tissue that would allow the assessment of organ-level response to drug-induced toxicity [11]. Jennifer Lewis’s group printed a range of cell densities from 0.1 to 10 million cells/mL to create 3D vascularized tissues and showed in vivo mimicry at high cell densities [12]. Researchers at University Medical Center Utrecht also explored cell densities from 3 to 10 million cells/mL but used 10 million cells/mL of human MSC and chondrocytes for in vivo implantation of osteochondral tissues [13]. Adam Feinberg also pointed out during the 3DHeals interview that his group was only able to get functioning cardiac heart muscle tissues when they started printing at very high cell densities at least 1-2 orders of magnitude from what most labs were printing at. This publication is pending but you can certainly refer to his interview here. As this industry matures, we are definitely observing higher cell densities being used to re-create in vivo like tissue constructs. Accessibility to higher cell volumes at a lower cost will also be important in fueling the rapid commercialization of tissue engineering and bioprinting applications.

As mentioned by the founder of RoosterBio, Jon Rowley, during the 3DHeals 2020 virtual conference Biofabrication Ecosystem panel discussion, cells should be envisioned as pieces of technology like a microchip. If a tissue engineering product developer has to figure out how to make their own microchip, it would take them many more years of development, time, and funding to get to their final product. It would be so much more beneficial and efficient to partner with companies who can provide a reliable and consistent cell supply chain to support their product development milestones. One thing for sure, the process innovation that we are witnessing today in MSC and cell therapy manufacturing is laying the groundwork for tissue engineering and bioprinting product commercialization success.

References:

[1] Charles Vacanti. The history of tissue engineering. J. Cell. Mol. Med. 2006. Vol 10(3), p569-576

[2] Wake Forest physician reports first human recipients of laboratory-grown organs. April 3, 2006. Press Release.

[3] Olsen et. al. Peak MSC – Are we there yet? Frontiers in Medicine. 2018. Vol 5: article 178.

[4] Izuta et.al. Meniscal repair using bone marrow-derived mesenchymal stem cells: experimental study using green fluorescent protein transgenic rats. The Knee. 2005. Vol 12, p217-223

[5] Dutton et.al. Enhancement of meniscal repair in the avascular zone using mesenchymal stem cells in a porcine model. J Bone Joint Surg. 2010. Vol 92(B): p169-75.

[6] Jung et.al. Enhanced Early Tissue regeneration after matrix-assisted autologous mesenchymal stem cell transplantation in full thickness chondral defects in a minipig model. Cell Transplantation. 2009. Vol 18, p923-932.

[7] Foldager et.al. Cell seeding densities in autologous chondrocyte implantation techniques for cartilage repair. Cartilage. 2012. Vol 3(2): p108-17.

[8] NCT03766217: Bone tissue engineering with dental pulp stem cells for alveolar cleft repair (CLOSE)

[9] NCT02748343: The clinical therapeutic effects and safety of tissue engineered bone

[10] Saris et.al. Matrix-applied characterized autologous cultured chondrocytes versus microfracture: two-year follow-up of a prospective randomized trial. Am J Sports Med. 2014. Vol 42(6): p1384-95

[11] Nguyen et.al. Bioprinted 3D primary liver tissues allow assessment of organ-level response to clinical drug induced toxicity in vitro. PLOS ONE. 2016. Vol 11(7): e0158674

[12] Kolesky et.al. Three-dimensional bioprinting of thick vascularized tissues. PNAS USA. 2016. Vol 113(12): p3179-84.

[13] Fedorovich et.al. Biofabrication of osteochondral tissue equivalents by printing topologically defined, cell-laden hydrogel scaffolds. Tissue Eng Part C Methods. 2012. Vol 18(1): 33-44

About the Author:

Mayasari Lim

Dr. Mayasari Lim is the West Coast Regional Account Manager for RoosterBio and an active contributor to the bioprinting community. She was the founder and CEO of SE3D, a startup focused on bringing bioprinting into the classroom to support future workforce development. Previously, she was an assistant professor in Bioengineering at Nanyang Technological University in Singapore. Her research expertise included stem cell bioprocess engineering, bioprinting, and regenerative medicine. She also mentors and teaches leadership and management courses at the Fung Institute for Engineering Leadership at UC Berkeley. Dr. Lim obtained her Ph.D. degree in Chemical Engineering at Imperial College London and her B.Sc. in Chemical Engineering at UC Berkeley.

Related Articles:

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Stem Cell Considerations for Bioprinting or Tissue Engineering

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Dr. Mayasari Lim Interviews upcoming 3DHEALS2020 speaker John O’Neil, Chief Scientific Officer of Xylyx (Podcast 🎙️)

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The post Meeting Cell Demands for Tissue Engineering and Bioprinting appeared first on 3DHeals.

About this Interview

Check out this informative interview between Dr. Mayasari Lim (Roosterbio/3DHEALS) and Adam Feinberg, Professor at Carnegie Mellon University, CTO and co-founder at FluidForm. Learn how Adam first got into 3D printing and bioprinting (years ago!), and how his team at CMU discovered the FRESH technique. Adam also shared his view on how to stay critical of convention and learn to think outside of the box. Adam will be speaking at the Organogenesis/Bioprinting panel at 3DHEALS2020 in June 2020. 

Podcast Transcript:

What inspired you to start your journey bio-fabrication/bio-printing?

Yes, I have been interested in 3D printing for a very long time. When I was a co-op student at Cornell, I did a co-op at Abiomed in the Boston area, working on total artificial hearts and at the time, rapid prototyping was used to make these mocks-up (in the late 90s) all in SLA but the resins were all phototoxic. I was enthralled by the idea of why can’t we just use this to build medical device directly, and why did we have to use this only to prototype and then as we moved to a device we need to test, we had to cast, machine it or mold it.

So, it was really clear what the potential was. Since then, 3D printing has also evolved from rapid prototyping to additive manufacturing entirely because the materials we were using and the systems in some degree have now progressed to the point where we can use what comes off the machine.

So since I was 20 years old, I have been fascinated by this technology. I started my lab in 2010 at Carnegie Mellon, this is when MakerBot appeared, FDM patent expired, the RepRap community started to grow and MakerBot is the company that really brought this to my attention. The first 3D printer we had was a Cupcake which is made out of laser cut wood in the lab, someone made a horrible extruder, perhaps not so at the time, but it was really poor that could just spit out a paste.

We were intrigued by the idea – we know we want to be able to additively manufacture some of the stuff, these technologies are becoming cheap and we had a Fab at Home, a project from Cornell at the time. I started the project at the time because EnvisionTec bioprinters at the time was $200,000 which at the time, I did not have. The idea of how we can bootstrap and the great thing about CMU is that we have engineering students everywhere.

So when I pitched the idea to students, they were ready to roll up their sleeves and get into it at CMU. Unlike many labs, we have been in this open-source bioprinter platform. All our publications have been based on open-source designs, mostly desktop grade FDM printers that we modified with custom extruders and turn them into bioprinters, that has been what we have used ever since. To me, it is exciting that we have been able to publish the paper last year in Science on a printer that we built under $1000, (I kinda like that).

There is so much innovation still in the space. If you can identify the problem that needs to be solve, very often it is the creativity and ingenuity that gets us the solutions and step changes, not necessarily a giant pile of cash. At least from an academic standpoint, and now we are trying to transition some of this work into a commercialized product in the bioprinting field, which is still very nascent compared to the rest of additive manufacturing which is a lot more mature. 

FRESH printing technique – how did you come up with this idea?

This was a project that was driven by my Masters then Ph.D. student, TJ Hinton. I challenged him to print hydrogels, there was not a great way to print at the time, they are soft and cannot support their own weight. We were kicking around different ideas and he had an epiphany about using gelatin microparticle support. For those who are not familiar, we basically print inside a gel, like a hair-gel material, we extrude inside of this material and whatever gets extruded directly into it, gets stuck in place. The reason the support bath we use is made of gelatin is that we can print at room temperature and then we just raise to body temperature so that the gelatin can melt away without destroying any parts of it.

He started this out with buying a packet of gelatin that you buy from the grocery store for baking which has a really rough powder. He basically put it into a dish, made it into a slurry, and printed into it and it kinda works. At first, when he showed it to me, he wasn’t too excited about it but when I saw it, I got super excited because I knew that we can fix the particles but this basic idea is going to work. That was probably 2012. We had our first paper in 2015 and another one last year and started our company in 2018. For a relatively simple idea to transition into commercialization, it is not a quick transition for sure.

Tell me more about your startup FluidForm – what is your vision?

Fluidform is focused on what FRESH enables, which is 3D printing of liquid materials. That is the unique thing that what we bring into the marketplace, by printing into this gel environment we can print a wide variety of fluids, some might gel and solidify right away, some might take days to cure. For example, you can print a silicon that you might normally mold but it will stay in a liquid shape for days until it adheres if that’s what you want. The vision of the company right now is focused on what we do best, which is in bioprinting of collagens and other hydrogels.

We are looking at this in 2 different ways – (1) the research market, which today there are already a lot of research grade bioprinters like Cellink and Allevi, and EnvisionTec, Regenhu and Advanced Solutions. We are looking to support all of the existing printers with our FRESH support materials, essentially to make them work better. Almost any extrusion based printer is compatible  with FRESH so if you go to Allevi’s or Cellink’s website, you will find LifeSupport. We also sell through Advanced BioMatrix, LifeSupport works really great with LifeInk, a bioink we use in our Science paper.

We do this because we hope that the research today is going to be applications for tomorrow. A lot of the tissue engineering and RM application are still early. Maybe they are in early clinical trial stage, but most are in animal studies but what we want to do is support that and hopefully provide better results with FRESH and people will become more successful. From a company’s standpoint we are hoping to look at other markets as well, and other areas of printing of collagen is of high value.

For example, in the area of tissue modeling and surgical planning, people are familiar with making models out of plastics from an MRI or CT scan. We are also making this out of a rubber like materials to give it a tissue feel but if we are print collagen then we can actually printing something that has the exact tissue feel. We see this as a potential interesting market, it’s not exactly TE or RM but it is using our core technology that can be used today. Also without worrying about regulatory challenges.

Can you share some of your early successes and failures in your work that change how you approach your work/research today?

Success/failure, to me, is part of the same thing. Research is really about learning from your failures and figuring out what the right path and success look like. In our Science paper, we had this 3D printed ventricle, and for the longest time, we thought that you have to mix the cells with your hydrogel bioinks cos that’s how everyone “bioprints” and that was really not giving us good results. Eventually, after hitting our heads against the wall many times, but we took a step back and looked at real tissue. For example, if you looked at muscle tissue, how it is organized? It has high-density collagen that makes fascia and other kinds of no-cell, just ECM that helps to organize muscle fibers. And muscles are just all cells, at very high concentrations, 1-2 orders of magnitude greater than what most people bioprint with.

So, we took that strategy and printed collagen by itself to define tissue compartments and changed our cell inks to 200-500,000 cells /mL (very high) and by doing that, we were finally able to get muscle constructs that we can print, function and beat. What was interesting is that this whole time we had MRI images thinking that we were mimicking nature, but we really were not, cos nature is not low cellular density hydrogels. We now applied this to a number of other tissue systems, not published yet but pretty robust as this is how most tissues are put together.

My advice to people is that, just because you read 50 papers and they are all doing it in a certain way, don’t think that’s the way it has to be done. There is so much space to innovate, we don’t yet know the answers, we are dealing with best guesses, we are trying things out, testing hypothesis. Sometimes it’s these little things that “let’s just do it this way” and then you realize that that does work, and then the next generation of things build on and it will hit the next roadblock and so on. Hopefully, that’s good advice!

Guest Biography: 

Professor Adam Feinberg is CTO and co-founder of FluidForm. The core technology of FRESH printing was developed in his Regenerative Biomaterials and Therapeutics lab at Carnegie Mellon University (CMU), where he is a Professor in the Departments of Biomedical Engineering and Materials Science and Engineering. His group develops materials-based, engineering strategies to control the self-organization and assembly of various cell types into tissues. Adam earned his Bachelor of Science in Materials Science and Engineering from Cornell University, and his MS and PhD in Biomedical Engineering from the University of Florida. He performed his postdoctoral work at the School of Engineering and Applied Science at Harvard University. He holds more than 20 US patents and patent applications, has authored over 45 publications, and is a member of the Materials Research Society, American Chemical Society, Society for Biomaterials, Biophysical Society, Biomedical Engineering Society, and the American Heart Association.

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The post Interview: Professor Adam Feinberg, Carnegie Mellon University, CTO and co-founder FluidForm appeared first on 3DHeals.

About this Interview:

An inspiration and candid conversation between Dr. Mayasari Lim and upcoming 3DHEALS2020 speaker John O’Neil, Chief Scientific Officer of Xylyx, a New York-based startup focusing on creating the perfect cell environment for a variety of biotech process including biofabrication/bioprinting, and tissue engineering. More specifically, Xylyx aims to translate the cell-specific extracellular matrix into biomaterial products with real-world impact. In this short interview, John shared with us his story of co-founding Xylyx, his vision for the future of bioprinting and tissue engineering, and lessons he learned from past successes and failures. John will be speaking with other startup founders and scientists on the Biofabrication Ecosystem panel on June 6th, 2020 at 3DHEALS2020. 

Guest Biography: 

Dr. John O’Neill is an expert on biomaterials and tissue- and disease-specific models aimed at transforming drug discovery. He has published extensively in the areas of extracellular matrix  (ECM), stem cell research, cell and tissue engineering, regenerative medicine, whole organ recovery and bioengineering, and the use of biomaterials in research, drug discovery and clinical applications. He is an inventor/co-inventor on over 10 issued and pending patents in biotechnology and imaging systems. Dr. O’Neill developed Xylyx Bio’s proprietary platform technology while completing his Ph.D. in Biomedical Engineering at Columbia University in the Laboratory for Stem Cells and Tissue Engineering under Gordana Vunjak-Novakovic, a world leader in tissue engineering, with expertise in regenerative medicine, translational stem cell applications, and disease modeling.

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The post Interview with John O’Neil, Chief Scientific Officer of Xylyx appeared first on 3DHeals.

Mr. Joshua Neubert has extensive experience managing entrepreneurial non-profit organizations, incentive prize competitions, educational programs, and startup enterprises. In 2012, he founded the Institute of Competition Sciences to create an online community and support system for academic competitions. His passions center on accelerating science, technology, and learning to make the world a more knowledgeable and inspiring place. Through the Institute of Competition Sciences, Mr. Neubert developed the Summit on Incentivized Innovation, launched the first collaborative community for challenge-based- learning, and helped develop and manage high-profile prizes providing over $8,000,000 in awards through partnerships with the Cleantech Open, Lemelson Foundation, NASA, Ideas42 and the Robinhood Foundation, Goodwill, Duke University, New Mexico State University, the Methuselah Foundation, the Actuarial Foundation, and others. Mr. Neubert has been a National Science Foundation Principal Investigator and has organized science, technology, engineering, and mathematics policy workshops with the NIH, HHS, NSF, NASA, and White House OSTP among others. Prior to founding ICS, Mr. Neubert helped NASA launch a $1.5 Million energy storage competition for advanced aerospace technologies. He previously led a team of over 500 volunteers and staff at the X PRIZE Foundation to create a STEM education program reaching 12,000 students. During this time, he developed the Spirit of Innovation Awards – a science and technology entrepreneurship competition for high school students – and later helped spin the program out as its own foundation built upon the legacy of Apollo 12 Commander Pete Conrad. He served as the founding Executive Director of the Conrad Foundation and launched the program into schools across the country reaching over 3000 students in its first three years. Mr. Neubert has designed and led grand challenges, hackathons, game jams, and other incentivized innovation programs for over 10 years and has served as Founder and CEO of the Institute of Competition Sciences since its founding in 2012.  He was trained as a Planetary Scientist at the Massachusetts Institute of Technology and continues to hold space science and exploration as a passion in his life. Mr. Neubert is a regular instructor, public speaker, and lecturer in entrepreneurship, innovation, and education topics particularly related to science and technology fields. Josh will be speaking at 3DHEALS2020.

Interview Video:

Maya: What inspired you to start your journey in biofabrication/bioprinting?

Joshua: I am not a biofabrication researcher myself, my career is more in the innovation generation space. I am a scientist by education but have turned my career to innovative project development and management. I love working on amazing things that have the potential to create really revolutionary changes. Bioengineering and particularly 3D bioprinting was introduced to me several years ago and I thought it was one of the most amazing opportunities to create change for good that I’ve ever seen.

Maya: Who inspired you the most along this journey in bio-printing/bio-fabrication?

Joshua: Anthony Atala’s work on bioprinting organs has been very inspiring to me. His ability to jump-start the whole field by demonstrating the transplantation of living, working organs (thin-walled like the bladder, trachea, etc…) was a revolutionary step. And his ability to tell the story about this was really inspiring.

Maya: Can you share some of your early successes and failures in your work that change how you approach your work/research today?

Joshua: Our work has been focused on creating new programs that help focus and incentivize the work in bioengineering to tackle critical challenges on the road towards ending the organ shortage through biofabrication of new tissues and organs. As we aren’t a research group ourselves, our successes and failures have been more around program development. For example, we started our effort in the New Organ Alliance by launching a $1M Liver Engineering Prize. We quickly realized that this prize was well ahead of its time. The hurdle we were asking to be solved was too far down the road. However, this led to the development of a “Roadmap to ending the Organ Shortage.” The roadmap identified 13 high-level challenges, and we were able to use this roadmap to bring NASA into our network to support our second prize, the Vascular Tissue Challenge which has been very successful in focusing research groups on the critical need to be able to create thick, vascularized tissues.

Maya: What is the biggest obstacle you have faced? If you have conquered them, what were your solutions?

Joshua: Funding. It seems that the research groups are there, they are ready to go, they are focused and interested in getting to the final destination, but the funding just isn’t there. We’re working on solving this, but it is still a major challenge. Our solutions have primarily focused on combining government and private funding, and focusing our attention on leveraging resources through incentivized innovation.

Maya: If you can go back 5 or 10 years, what would you do differently?

Joshua: Do the roadmap first before jumping into the Liver Prize. Focus early on the development and promotion of the critical hurdles ahead in the bioengineering space. This has really helped us focus our work and bring others together around common challenges.

Maya: What advice would you give to a smart driven college student in the “real world”? What bad advice you heard should they ignore?

Joshua: Talk to people. Don’t just sit in the lab. Work with your advisor to connect with others and build your network in the field. Learn from many mentors and think about what role your research could play in a bigger picture that the whole industry is interested in.

Maya: What is the biggest challenge you are facing in 2020 and how do you plan to overcome this?

Joshua: Funding. The coronavirus has made it extremely difficult to keep programs like ours growing. We are continuing to work with government partners and new philanthropic groups to explore our next steps.

Maya: What was/is the biggest risk you took in your career?

Joshua: Left my Ph.D. research to join a non-profit organization focused on incentivized innovation. This changed the whole trajectory of my career.

Maya: What do you enjoy in your spare time? What are you passionate about outside of your work/3d printing?

Joshua: Space exploration and science fiction, and gaming. I love fantasy and sci-fi games, books, and movies. I’d love to actually go to the moon or mars one day.

Maya: What gets you up in the morning every day? 

Joshua: My alarm (I also like bad jokes). But really, I get excited every day about doing things that matter to other people. It’s a bit narcissistic that I like having other people like me, and like the things, I’m doing, but I think I’m doing really cool things. And so long as I can keep working on programs that make a difference for people, that gets me going in the morning.

Maya: What does the word “3DHEALS” mean to you?  =)

Joshua: I like that it has such a breadth to it. There is so much that can be done with “3D” and “Heals” it isn’t just about bioprinting but is about all the other ways that it can be used in healthcare. This multi-disciplinary connection around a common theme really seems to bring a great group of people together.

About the Interviewer:

Mayasari Lim is the West Coast Regional Account Manager for RoosterBio and an active contributor to the bioprinting community. She was the founder and CEO of SE3D, a startup focused on bringing bioprinting into the classroom to support future workforce development. Previously, she was an assistant professor in Bioengineering at Nanyang Technological University in Singapore. Her research expertise included stem cell bioprocess engineering, bioprinting, and regenerative medicine. She also mentors and teaches leadership and management courses at the Fung Institute for Engineering Leadership at UC Berkeley. Dr. Lim obtained her Ph.D. degree in Chemical Engineering at Imperial College London and her B.Sc. in Chemical Engineering at UC Berkeley. She is 3DHEALS Bioprinting Ambassador as well as a speaker and moderator for 3DHEALS2020.

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The post Interview with Joshua Neubert, NASA Vascular Tissue Challenge (Video) appeared first on 3DHeals.

Stem cells no doubt play an important role in many tissue engineering and bioprinting applications. While the choice of stem cells is very much application dependent, and often requiring more than one cell type, mesenchymal stem/stromal cells (MSCs) has served as the workhorse for numerous tissue engineering and bioprinting applications. Most commonly, MSCs are used in bioprinting of bone [1-3] and cartilage tissue [4, 5], but they are also used in the skin [6], cardiac, and wound healing [7] applications often in combination with other cell types that may leverage the regenerative properties of MSCs. Moreover, MSCs already have a proven safety profile, there are over 1000 MSC clinical trials worldwide to date. This makes MSCs an attractive cellular starting material for creating more sophisticated tissue-engineered products and future organs supported by fabrication technologies such as bioprinting.

The first important consideration when designing or developing a tissue-engineered product is the selection and characterization of cellular starting material. The development of a patient-specific product, as opposed to an off-the-shelf product, will require different CMC (Chemistry, Manufacturing, and Controls) strategies. Donor criteria and eligibility will differ slightly depending on whether it is for autologous or allogeneic use. The criteria for autologous use are generally less stringent than those for allogeneic use, cells from allogeneic sources must pass all infectious disease testing and be able to withstand longer storage conditions. Donor eligibility guidelines from the FDA are provided under regulations at 21 CFR Part 1271. Pertinent to MSCs, the choice of tissue source, whether it is from the bone marrow, adipose tissue or umbilical cord will depend on the target use. Characteristics of MSCs are notably different from different sources [8], their differentiation potential and thus potential clinical applications can also vary [9]. For allogeneic cells, donor variability is inherent, so it is critical to determine suitable methods and assays to characterize the cells in order to ensure consistency of performance. The most basic characteristics would include cell enumeration, viability, and phenotypic assays, but other methods such as biological activity and functional assays should also be included.

In addition to the need for high quality and consistent cell source, tissue-engineered products often require high quantity or volumes of cells. As an example, in the bioprinting of tissue constructs such as bone, the challenge here is no longer the choice of biomaterial or 3D design and technology necessary to create the desired scaffold but it is in generating enough starting cellular material for printing a biologically relevant tissue for transplant. The first group that attempted to do this and 3D bioprinted the superior half of an adult human femur was John Fischer’s lab [2], this print consumed 720M human MSCs (hMSCs) to seed this bone scaffold. The number of cells used in this single bioprint is at least 10-fold more than a typical tissue engineering experiment performed in an academic lab. At the time, this construct was the first successful demonstration of a large bioprinted bone tissue construct (20x larger than average) that achieved high viability throughout the graft and increased expression of osteogenic markers. An impressive accomplishment indeed!

In order to accelerate this into the clinic, a full femur replacement would require 1.5B hMSCs as a starting cellular material. These cells will not only need to be manufactured under current Good Manufacturing Practice (cGMP) to meet stringent regulatory requirements but also be well-characterized, tracked, and used at the appropriate cellular age by assessing their population doubling levels. The next step is to determine appropriate cell lot sizes required at each stage of the development process from product development to clinical manufacturing all the way through commercial manufacturing. These are important factors to consider as early as possible in the product and process development stage as this is commonly neglected – the process and its scalability. Many people make the mistake of going into clinical trials with sub-optimal processes and without a full understanding of their process, resulting in more painful consequences that may or may not be recoverable. Early and thoughtful planning is always the first step in making sure that you are setting yourself up for success. As Stephen Covey states as his second habit in The 7 Habits of Highly Effective People, “Begin with the End in Mind”.

References:

[1] Kolesky DB, Homan KA, Skylar-Scott MA, Lewis JA. Bioprinting thick vascularized tissues. Proceedings of the National Academy of Sciences Mar 2016, 113 (12) 3179-3184; DOI:10.1073/pnas.1521342113

[2] Nguyen BNB, Ko H, Moriarty RA, Etheridge JM, Fisher JP. Dynamic bioreactor culture of high volume engineered bone tissue. Tissue Eng Part A (2016) 22:263–71. doi: 10.1089/ten.tea.2015.0395

[3] Du M, Chen B, Meng Q, Liu S, Zheng X, Zhang C, Wang H, Li H, Wang N, Dai J. 3D bioprinting of BMSC-laden methacyrlamide gelatin scaffolds with CBD-CMP2-collagen microfibers. Biofabrication (2015) 7: 04410

[4] Möller T, Amoroso M, Hägg D, Brantsing C, Rotter N, Apelgren P, Lindahl A, Kölby L, Gatenholm P. In Vivo Chondrogenesis in 3D Bioprinted Human Cell-laden Hydrogel Constructs. Plast Reconstr Surg Glob Open. 2017 Feb 15;5(2):e1227. doi: 10.1097/GOX.0000000000001227. PubMed PMID: 28280669; PubMed Central PMCID: PMC5340484.

[5] Apelgran P, Amoroso M, Lindahl A, Brantsing C, Rotter N, Gatenholm P, Kolby L. Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo. PLoS One. 2017 Dec 13;12(12):e0189428. doi: 10.1371/journal.pone.0189428

[6] Wang X, Li C, Zheng Y, Xia W, Yu Y, Ma X. Bone marrow mesenchymal stem cells increase skin regeneration efficiency in skin and soft tissue expansion, Expert Opinion on Biological Therapy (2012),12:9, 1129-1139, DOI: 10.1517/14712598.2012.704016

[7] Skardal, A. , Mack, D. , Kapetanovic, E. , Atala, A. , Jackson, J. D., Yoo, J. and Soker, S. (2012), Bioprinted Amniotic Fluid‐Derived Stem Cells Accelerate Healing of Large Skin Wounds. STEM CELLS Translational Medicine, 1: 792-802. doi:10.5966/sctm.2012-0088

[8] Hass R, Kasper C, Böhm S, Jacobs R. Different populations and sources of human mesenchymal stem cells (MSC): A comparison of adult and neonatal tissue-derived MSC. Cell Commun Signal. 2011;9:12. Published 2011 May 14. doi:10.1186/1478-811X-9-12

[9] Berebichez-Fridman R, Montero-Olvera PR. Sources and Clinical Applications of Mesenchymal Stem Cells: State-of-the-art review. Sultan Qaboos Univ Med J. 2018;18(3):e264–e277. doi:10.18295/squmj.2018.18.03.002

About the Author:

Mayasari Lim

Dr. Mayasari Lim is the West Coast Regional Account Manager for RoosterBio and an active contributor to the bioprinting community. She was the founder and CEO of SE3D, a startup focused on bringing bioprinting into the classroom to support future workforce development. Previously, she was an assistant professor in Bioengineering at Nanyang Technological University in Singapore. Her research expertise included stem cell bioprocess engineering, bioprinting, and regenerative medicine. She also mentors and teaches leadership and management courses at the Fung Institute for Engineering Leadership at UC Berkeley. Dr. Lim obtained her Ph.D. degree in Chemical Engineering at Imperial College London and her B.Sc. in Chemical Engineering at UC Berkeley.

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The post Stem Cell Considerations for Bioprinting or Tissue Engineering appeared first on 3DHeals.

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Nothing excites me more than to see the progress we have made in the world of bioprinting. It really hasn’t been that long ago when the first idea of bioprinted organs came about, since then many research groups have jumped onto the bandwagon of bioprinting, and in the last 5 years, we have witnessed the burst of bioprinting companies that spun out of academic labs. To evaluate where we are today, here’s a quick recap of the progress made in 2018, by companies and research institutes propelling the bioprinting industry. 

On research innovations, several companies announced key scientific milestones that are fulfilling the promises of bioprinting. In April 2018, Organovo announced key breakthrough developments in their bioprinted liver tissue models for evaluating a spectrum of liver diseases and intestinal tissue models. This breakthrough is allowing researchers to develop new drugs for non-alcoholic fatty liver tissue diseases (NAFLD) more effectively at early stage development. One of the newest bioprinting startups, Biolife4D, announced their success in bioprinting a human cardiac patch, achieving a major milestone demonstrating their ability to 3D print human cardiac tissue derived from induced pluripotent stem cell (iPSC) technology. Our local Bay Area resident bioprinting company, Prellis Biologics, co-founded by two female scientists, announced the first pre-vascularized tissue scaffolds, Tissue Blanks TM designed for building complex 3D cell and tissue constructs. This is another major milestone which has, in the past, prohibited researchers to build larger constructs due to the lack of vasculature.

On the technology side, more innovative platforms are being developed. Poietis, a French-based bioprinting company, announced the launch of their next generation laser-assisted 4D bioprinting system. Aether, another Bay Area resident company, announced their new AI-powered 3D medical imaging software designed to enhance bioprinting capabilities, especially for organ printing. Allevi partnered up with Made in Space to create the first zero-gravity tissue 3D printer (Allevi ZeroG) and Cellink announced their partnership with Prellis Biologics to create the first holograph-based volumetric 3D bioprinter, Holograph-X. Both equally exciting developments in the bioprinting world to be looking forward to in perhaps 2019. 

The year of 2018 was also a year where more research collaborations are being made. Aspect Biosystems announced several academic collaborations with various universities including a project to 3D-bioprint skin with Dalhouise University. In addition, they also announced a collaboration with JSR corporation to create liver tissue. Poietis also partnered with Prometheus to develop high precision 3D bioprinting of tissue engineered advanced therapeutic medical products for skeletal regeneration. The Advanced Materials + BioEngineering Research (AMBER) center headquartered in Dublin has partnered with JnJ to tackle osteoarthritis by bioprinting implants that regenerate. 

On the funding side, the Food and Drug Administration (FDA) issued $2.6M in grants toward 3D bioprinting and biomedical research last year. Awardees included Harvard University, Carnegie-Mellon University, Rutgers University, Georgia Institute of Technology and Massachusetts Institute of Technology. Oxford spin-out bioprinting company, Oxsybio raised $10M GBP (~$14M USD) in Series A funding last year. Biolife4D initiated their Regulation A+ equity crowdfund investment offering in February 2018, this round is expected to close end of January 2019. 

With so much progress being made already in 2018, I cannot wait to see what lies ahead for us in this coming year. Companies and academics in the field are increasing collaborative efforts to bring together a convergence of technologies and knowledge that is only going to help us make in-roads and continued progress in healthcare and regenerative medicine.

About the author:

Mayasari Lim

Dr. Mayasari Lim is the West Coast Regional Account Manager for RoosterBio and an active contributor to the bioprinting community. She was founder and CEO of SE3D, a startup focused on bringing bioprinting into the classroom to support future workforce development. Previously, she was an assistant professor in Bioengineering at Nanyang Technological University in Singapore. Her research expertise included stem cell bioprocess engineering, bioprinting, and regenerative medicine. She also mentors and teaches leadership and management courses at the Fung Institute for Engineering Leadership at UC Berkeley. Dr. Lim obtained her Ph.D. degree in Chemical Engineering at Imperial College London and her B.Sc. in Chemical Engineering at UC Berkeley.

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The post How Far Ahead: A Look Back at 2018 in the World of Bioprinting appeared first on 3DHeals.