The convergence of 3D assembly technologies such as 3D printing and electrospinning with cell therapy advancements is allowing researchers an unprecedented look into the mechanics of cellular behavior to study the development of diseases such as cancer.
By allowing the construction of three dimensional model systems, we are now able to look into the behavior of cells at the microscopic level by placing cells in synthetic environments that more closely resemble their native ones.
A key to constructing such 3D systems is the use of biological matrix proteins including fibronectin, vitronectin, laminin, collagen and elastin.
A new study by Dr. Wei Sun’s lab at Drexel University is further proof of this.
In the study, titled “Three-dimensional printing of Hela cells for cervical tumor model in vitro,” the authors 3D printed a tumor-like structure with gelatin, fibrous proteins including fibronectin, and cervical cancer cells layer by layer.
This generated a structure that resembled the fibrous proteins that make up the extracellular matrix of a tumor.
Cell proliferation was compared between the 3D model system and a regular 2D plate system. The results showed that Hela cells showed a higher proliferation rate in the 3D printed system where they formed cellular spheroids, but formed monolayer cell sheets in 2D culture. Moreover, a cell viability of over 90% was observed using the 3D printing process.
The study, published in Biofabrication, can be accessed here.
Another recent study, titled “Regulators of Metastasis Modulate the Migratory Response to Cell Contact under Spatial Confinement” and published by Dr. Anand Ashtagiri’s lab at Northeastern University focused on studying the migration and development of cancer cells.
To study this, they also used a model ECM system with fibronectin, which provided a substrate for cell migration.
The study highlights a characteristic fibrillar dimension at which effective sliding was achieved.
Both studies are examples of the critical role ECM proteins play in the development of model systems to study cell-based disease modeling.
Understanding how these ECM proteins work is the key to studying how they can be used in modeling cell migration and disease development. A supplier like Akron possesses a wide knowledge base of expertise that includes not only sourcing of these proteins, but also their mode of action and the ways in which they can be implemented in bioassays and 3D systems.
Options like viral inactivation – an Akron industry first – further enhance the clinical utility of these proteins. Contact us to discuss.
A remarkable new step toward heart regeneration: a beating 3D micro heart from induced pluripotent stem cells.
This is what a new study by Dr. Bruce Conklin’s lab at the Gladstone Institute of Cardiovascular Disease and the University of California, San Francisco described in a new manuscript published in the journal Scientific Reports.
Engineering micro heart muscle from induced pluripotent stem cells has been challenging: obtaining aligned and unidirectionally contracting tissue without requiring unfeasibly high numbers of cells and requiring easily manufactured materials has been a traditional constraint that has limited progress in this area. Moreover, differentiating iPS cells into heart cells produces weak and immature heart cells, making working with these cells difficult.
The new study overcame these constraints by creating a system wherein the authors seeded a mixture of cardiomyocytes and fibroblasts derived from iPS cells into stencils containing – in the authors’ own words – “dogbone” through-holes comprised of a high-aspect ratio “shaft” flanked on either end by square “knobs.”
To clarify this design, see image in Panel A below.
Such confinement of cardiomyocytes and fibroblasts into rectangular canals aligned in three dimensions and induced uniaxial beating within these canals.
Moreover, such alignment allowed the authors to work with significantly fewer cells than traditionally needed for engineered micro heart tissue systems.
This is a remarkably simple solution to a complex problem.
These cells were also cultured for a short time, during which they adopted an in vivo-like morphology, and exhibited some functional aspects of adult cardiac tissue. These paramters, however, require further investigation and experimentation.
The authors intend to follow this up with more advanced systems that allow parallelization and automation, to try and achieve a higher throughput screening system of artificial mini hearts.
See video below of the beating mini hearts in action.
And though more work is necessary to establish whether this iPS-CM-based micro heart system can accurately model adult heart tissue, it is an interesting example of how geometry and environment can regulate the behavior of cells and tissue towards fully functional organs.
On the heels of Akron Biotech’s acceptance of our Drug Master File for interleukin-2 by the FDA, it is appropriate to highlight what such an acceptance means and why one should be concerned with the quality of any interleukin-2 used in their immunotherapy projects.
As the first immunotherapy approved by the FDA for treatment of metastatic melanoma, interleukin-2 (IL-2) has come a long way from where it was two decades ago: the subjects of extensive research, it remains one of the most important cytokines used in the development of cancer-targeting immunotherapies. There is compelling data of immune activation leading to cancer regression after treatment with IL-2 activated T-cells, and such research continues.
But not all interleukin-2s are the same: the path to the clinic – and eventually, regulatory approval and commercialization – is paved with a number of checkpoints that are often overlooked.
Oftentimes, when it comes to sourcing raw materials – including interleukin-2 – one of the most important factors taken into consideration is cost. This is a reasonable selection criterion, but one that fails to address the complexity and severity of proper selection of IL-2.
No consideration of cost should come at the expense of quality – but what does quality mean, exactly?
Across the industry, there is widespread confusion about terms such as “GMP,” “research grade,” “injectable grade,” “clinical grade,” as well as the importance and use of batch records, drug master files, SOPs and quality documentation. Common questions one should be expected to know the answers to are: when are these documents needed? Who is responsible for writing them? What kind of data and information is needed for each of them?
If you were asked which of the above are required for pre-clinical development of an IL-2-based immunotherapy, would you know? What about clinical trials?
Moreover, if you are currently a user of interleukin-2 either as a raw material in a current immunotherapy development process or as a reseller to end users , are you aware of the type of documentation available for the IL-2 and how the needs for this documentation change as the development pipeline progresses? What is needed at the research and development phase is very different to what one needs when submitting documents to the FDA. Not being adequately prepared often causes scrambles down the road and, more worringly, regulatory denials.
This happens often and has caused regulatory issues that have costed companies significantly more than it would have cost them had they sourced their raw materials correctly.
It is very common to underestimate the need for such documentation as well as background data supporting these products, but most of all, misunderstand the need for continued support and updates to such documentation.
Unfortunately, many IL-2 suppliers also do not have a full understanding of the clinical development process and as such, do not translate this information adequately to customers thereby further promoting miseducation about this.
Just like not all immunotherapies are the same, not all IL-2s are the same either: ask yourself, what kind of IL-2 is your supplier providing? What kind of data – stability, functionality, pre-clinical and clinical – are they able to provide you?
Do not be surprised if most of this information is not available, or if you are not able to obtain answers to such questions.
Make sure that your supplier is able to provide a full portfolio of information, data, but most importantly, support to help guide the development process.
While this post should not come across as a direct promotion of Akron Biotech’s interleukin portfolio, we want to highlight the unique prospect of working with us on your immunotherapy products: not only is our interleukin-2 (and many other growth factors) manufactured as pharmaceutical grade, the product is supported by an extensive dossier of data – from stability, to functionality, all based on international standards – that allows us to provide the right product for the right stage of development.
More importantly, the flexibility of adjusting documentation as your process changes is possible and done easily and quickly.
Ask yourself: is your current interleukin-2 right for you?
In an Advanced Manufacturing report for April 2016, A Snapshot of Priority Technology Areas Across the Federal Government, the US Federal Government’s Subcommittee for Advanced Manufacturing of the National Science and Technology Council highlighted areas of US manufacturing technology areas of emergeing priority.
Among them, regenerative medicine was highlighted as one of the areas of interest for continued future financial support.
The Federal Government invested $2.89 billion between 2012-2014 on regenerative medicine, money which primarily went to the development of therapeutic techniques and technologies.
Current funding programs developing new regenerative medicine products and technologies come from DoD, Food and Drug Administration (FDA), NIH, NIST, DOE National Nuclear Security Administration, NSF, and the Department of Veterans Affairs (VA).
Some of the investment programs supported by these agencies include the FDA’s Mesenchymal Stem Cell Consortium, DoD DARPA’s various macrophysiological systems programs, HHS NIH NHLBI’S Stem Cell-Derived Blood Products for Therapeutic Use Program and more.
The report highlights that future support for regenerative medicine manufacturing technologies will require collaborative work. Specifically, the report lists some expectations from teams developing future technologies which will lead to more efficient products. These include, quoted directly from the report:
To manufacture specific tissues, a project team must have advanced engineering skills combined with a deep understanding of the physiology of those tissues and the organ systems in which those tissues function
Controlling and characterizing the biologic activity through discovery, triggers, and terminations of the appropriate effector and control mechanisms at the individual cell and population levels will require the expertise of genomic scientists
Efficiencies at each step are essential to keep costs and raw materials usage down, and to keep fragile products (i.e., cells, tissues) viable, while a constant focus on usability will ultimately help enable adoptability.
Biomanufacturing for Regenerative Medicine is also listed as one of the areas of future investment for which the Government has issued a Request for Information to create a Manufacturing Innovation Institute.
Encouraging interest from the US Government in supporting further development in regenerative medicine highlights the need for more innovation in new products, technologies and strategies for cost reduction in regenerative medicine.
Akron is pleased to announce acceptance by the Food and Drug Administration (FDA) of a Drug Master File (DMF) for our immunotherapy cytokine interleukin-2.
We invite all customers interested in cross-referencing our DMF for IL-2 in one of your submissions to government or regulatory agencies the FDA to contact us.
A few weeks ago, we wrote about Akron’s commitment to supporting immunotherapy advances through our capabilities in product and technology development which includes our sphisticated network of cytokines for research and clinical grade.
At the forefront of those is interleukin-2, which is fully documented and backed by an extensive dossier of data including extended stability and functionality documentation. You can get more information on our GMP-grade IL-2 here.
Now, we are pleased to announce the submission and acceptance by the FDA of a Drug Master File for Interleukin-2.
Drug Master Files are confidential documents submitted to the FDA which outline, in detail, manufacturing and procedural details for drugs or biological materials used in animals or humans.
DMFs were extensively described in one of our recent blog posts, and we invite you to read it here.
If you are interested in requesting a letter of authorization to reference our DMFs, please get in touch with us so that we may provide such authorization.
With IL-2 at the forefront, Akron also supplies a variety of other growth factors the roles of which are to enhance immunological response to clients that range from clinical-stage entities to research and development labs and academic institutions.
The acceptance of this DMF further confirms Akron’s continued commitment to quality and providing the highest level of support and transparency to our immunotherapy clients.
New Seminar on FDA’s Regulations for Stem Cell Therapy Development
If you are interested in learning more about the FDA’s current guidelines for the approval of stem cell therapies as well as the different guidances regulating to the various approval processes (INDs, BLAs etc.), then a new seminar might be of significant benefit.
FDA’s Regulation of Regenerative Medicine including Stem Cell Treatments, Tissue Engineering and Gene Therapies is a two-day seminar aimed at those involved, in some way, in the cell therapy development process – from regulatory, quality and compliance personnel to clinicians and researchers – that aims to present in detail the various guidances associated with the drug development and approval processes, but also design, GMP and GLP guidances, as well as product labeling and marketing.
The seminar will take place over two sessions: the first one is in Irvine, CA on July 14-15, and the second is in Newark, NJ from October 15-16, 2016.
The speaker at the seminars is Dr. Thomas J. Webster, Department Chair and Professor of Chemical Engineering at Northeastern University.
For more information and to register, follow this link.
FDA Postpones Public Hearing on Draft Guidances and Adds Scientific Workshop
The FDA was to hold a public hearing for interested stakeholders to provide feedback on their recently-released Draft Guidances Relating to the Regulation of Human Cells, Tissues or Cellular or Tissue-Based Products. The hearing was to take place on April 13, 2016, at the FDA’s White Oak Campus in Silver Spring, Maryland.
The purpose for the hearing is to provide stakeholders an opportunity to discuss the FDA’s proposed regulation of HCT/Ps regarding homologous use, same surgical procedure exception, minimal manipulation, and adipose tissue.
Due to the high interest, the FDA has postponed the hearing to a date later this year.
However, ina ddition to the hearing, the FDA now plans to also add a scientific workshop to “gather information from manufacturers of cell based products, clinical researchers […] regarding the generation of scientific evidence to facilitate the development of safe and effective cell based therapeutics.”
More details on the hearing and workshop will be provided once known.
Bipartisan Policy Group Releases Cell Therapy Document
This Winter, the Bipartisan Policy Group (BPC) – a Washington, DC-based political think-tank promoting various agendas aimed at advancing areas of national interest –
unveiled a document, titled Advancing Regenerative Cellular Therapies, which aims to presents new pathways for the development of cellular therapies that would allow for the more rapid and clinically efficient translation of discoveries to viable commercial therapies.
The document proposes a number of significant recommendations to the FDA. The bulk of the new proposals are centered around a “conditional-approval” clause. In summary, their recommendations are:
• A proposal for the FDA to grant a time-limited, conditional approval for cell-based therapies based on preliminary clinical evidence of safety and efficacy, without Phase 3 trials;
• Allow for patients to be granted limited access to these conditionally-approved therapies
• Require the sponsor of the conditionally-approved therapies to submit a BLA within three years of receiving conditional approval or negotiate with the FDA an extended conditional period to collect additional data
• Permit reimbursement during the conditional approval period
The entire document can be accessed here.
These proposals have been met with caution, and they have been called misguided and controversial elsewhere – particularly with respect to the “conditional approval” clause.
We will not comment on the validity of these proposals, but invite you to read the entire report and look for the industry’s reactions directly.
Culturing stem cells on substrates that are fabricated in three dimensions provides important advantages to their therapeutic potential. 3D scaffolds have thus emerged as powerful substrate systems for regenerative medicine applications. Significant advances in the technologies used to generate these systems have emerged, and some have even reached commercial status.
A new just-published study further expands the potential of 3D scaffolds in the treatment of CNS disorders.
Dr. Prabhas Moghe, Professor at the Department of Chemical and Biochemical Engineering at Rutgers University, and colleagues have developed a 3D micro-scaffold platform that promotes reprogramming of stem cells into neurons, and supports growth of neuronal connections capable of transmitting electrical signals.
Titled Generation and transplantation of reprogrammed human neurons in the brain using 3D microtopographic scaffolds, the study was published this week in the journal Nature.
The scaffolds are based on electrospun nanofibers – a technology that involves spinning long, thin fibers of various synthetic or natural materials into custom-aligned shapes.
The authors used aelectrospinning to generate fibrous substrates, while spin coating was used to create 2D polymer film controls from tyrosine-derived polycarbonate pDTEc.
The assumption was that such 3D microstructures would support neuronal networks that exhibit improved levels of retention and engraftment following transplantation into the brain.
Through a combination of scaffold geometry and media supplementation, the authors demonstrated suppression of proliferation of undifferentiated iPSCs.
After demonstrating successful engrafment of induced neuronal cells into the CNS in an ex vivo organotypic brain slice model, the authors followed this with an in vivo demonstration of scaffold functionality.
Transplantation of induced neuronal cells in 3D scaffolds into the mouse striatum showed that the percentage of viable cells after 3 weeks was an order-of-magnitude larger than that obtained with isolated single cells.
The fiber material, the thickness of the fibers, their porosity and the scaffold geometry were identified as important factors that together play a critical role in the functionality of these scaffolds compared to standard 2D fibrous substrates.
More questions about electrospinning? Need a custom scaffold system? Contact us here.