stem cells

cGMP Compliance: What Does It Mean for Your Cell Lines?

As the promise for cell-based therapy grows, the interest in making clinically relevant cell lines has skyrocketed for industrial and academic researchers alike. For translation into human therapies, cell-based products must be made following current Good Manufacturing Practice (cGMP). Many groups have already claimed to generate cell lines that are “cGMP-compliant,” “cGMP-ready,” or “certifiable under cGMP.” But what does it take to be truly cGMP-compliant, and what practices can you introduce in your lab to comply with cGMP standards?

A common misconception in the United States is that a facility is granted a ‘cGMP license’ from the government to manufacture cGMP-grade products. Rather, the Food and Drug Administration (FDA) evaluates the manufacturing process for each product to determine if it is compliant with cGMP standards. The primary concern when it comes to deriving cell-based products for therapies is making sure that the product is derived in a safe and reproducible manner. To ensure maximum quality assurance, researchers should

• choose reliable, xenogeneic-free raw materials,
• establish and monitor a clean environment,
• qualify all equipment and software,
• remove variation in laboratory procedures by creating detailed Standard Operating Procedures (SOPs) and by providing rigid process validation at each step.

Nevertheless, even establishing robust quality assurance does not imply that the process is scalable for commercial production. In the world of biologics, “the product is the process.” A requisite step to ensure a smooth transition to cGMP practice is to ensure that the process of manufacturing is not altered due to changes in production scale. For example, depending on the therapy, millions or billions of cells may be required for a single patient. Therefore, it is in the best interest of the researchers to develop a scalable method at the beginning to avoid revamping the entire process (e.g., changing from adherent cells to suspension). Along these lines, the quality control (QC) requirements of cell-based products should be carefully considered and not have to include difficult-to-assay tests. For example, some cell lines have been qualified as cGMP-compliant upon conversion from research-grade conditions to cGMP quality standards. Rigorous tests were performed on the converted lines to ensure that the cells were free of contamination. Even though strict measures were carried out to ensure cGMP compliancy, deriving cell lines in this manner makes scalability and reproducibility a challenge. Ideally, the entire process of deriving cell products for clinical use should be performed under cGMP conditions: from the acquisition of human tissue to the manufacturing, testing, and storage of derivative cell products.

Another important consideration when instituting cGMP-compliance is documentation. Each process must be described with rigorous SOPs, the training of individual manufacturing operators must be well-documented, and the entire established process must be validated and well noted. Failure to document—in the eyes of the FDA—is often equated with failure to perform the underlying activity. It is equally important to remain ‘current.’ The FDA expects manufacturing processes to stay up-to-date with current regulations, even as policies change.

For an academic lab, closely aligning with cGMP standards can ensure that the resulting cell lines are comparable to other truly cGMP-produced products used during clinical trials. It is in the best interest of academic researchers to establish rigorous SOPs and use qualified reagents and equipment, even if it is not possible to carry out all steps in a certified cleanroom. Whenever possible, it is advisable to acquire truly cGMP cell lines from appropriate sources for preclinical projects; if prohibited by costs or other reasons, it is recommended to use a protocol that is as close to cGMP as possible.

Tags: , , , , ,

New Allele Biotech Publication on Stem Cells

Feeder-Free Reprogramming of Human Fibroblasts with Messenger RNA
Current Protocols in Stem Cell Biology • November 13, 2013
DOI: 10.1002/9780470151808.sc04a06s27

Authors: Luigi Warren, Jiwu Wang

This unit describes a feeder-free protocol for deriving induced pluripotent stem cells (iPSCs) from human fibroblasts by transfection of synthetic mRNA. The reprogramming of somatic cells requires transient expression of a set of transcription factors that collectively activate an endogenous gene regulatory network specifying the pluripotent phenotype. The necessary ectopic factor expression was first effected using retroviruses; however, as viral integration into the genome is problematic for cell therapy applications, the use of footprint-free vectors such as mRNA is increasingly preferred. Strong points of the mRNA approach include high efficiency, rapid kinetics, and obviation of a clean-up phase to purge the vector. Still, the method is relatively laborious and has, up to now, involved the use of feeder cells, which brings drawbacks including poor applicability to clinically oriented iPSC derivation. Using the methods described here, mRNA reprogramming can be performed without feeders at much-reduced labor and material costs relative to established protocols.

Allele iPSC Service and Technology Licensing Contact: http://www.allelebiotech.com/cell-line-and-culture-services/#ips-line

New Allele Product of the Month: FP-nAb™ products for 100% pull-down

Tags: , , , , , , , , , , , ,

Wednesday, November 13th, 2013 iPSCs and other stem cells No Comments

Allele Biotech Receives $200,000 Grant to Update Its mRNA Reprogramming Commercial Products and Services

On June 10, 2013 Allele received an SBIR award from the National Institute of Drug Abuse (NIDA/NIH) entitled “Revolutionary Technology for Efficient Derivation of Human iPSCs with Messenger RNA”. The goal of the proposed project is to provide to the biomedical research market an advanced reagent kit and services for highly efficient reprogramming of high quality human induced pluripotent stem cells (iPSCs). At the core of this kit is the Allele team’s recent development transcribed messenger RNA (mRNA). Compared to other reprogramming methods, such as lentivirus, Sendai virus, protein, small molecules or any combinations of these reagents, our new generation of the mRNA method often requires less than half the time while sometimes achieving “bulk conversion” efficiency.

While the Allele reprogramming technology was designed for clinical use as the process is feeder-free, xeno-free, chromosome integration-free, as well as without the need for cell splitting, PI, Dr. Jiwu Wang states, “Our purpose of executing the NIH-funded research it to make our method so easy that any researcher can integrate iPSC into his or her projects.” In addition to the extremely high efficiency, mRNA-generated iPSCs should also be more stable because there are no genetic alterations, more uniform among all clones as there is no clonal event, and ultimately suitable for future autologous cell therapy now that creating iPSCs from patient tissue cells should no longer be the rate-limiting steps.

Allele’s business model is to provide cGMP-grade iPSCs to pharmaceutical companies and perform large scale reprogramming by partnering first with university-affiliated hospitals. Great progress has been made in both directions, which has prompted the initiation of a cGMP unit within Allele’s newly acquired building in San Diego.

Tags: , , , , , , , ,

Solving the Big Problems of the World

Science, by nature, is something you do without knowing for sure that it will work. By doing an experiment, testing a theory, or tabulating large data sets to find statistical significance, researchers make small discoveries or incremental improvements on technologies. It is easy for any researcher to get buried under the enormous amount of experimental details while trying to complete a project that lasts for months and years. For a team or an organization, however, it is critical to create a level of alertness of the big questions we try to answer – why are we doing this line of research? Is the technology or theory being developed going to be disruptive in terms of changing the ways of thinking in its field or solving a big challenge that faces the world?

The world does not lack for challenges: there may not be any ice left at the North Pole as early as 2015, there are still a billion people who need reliable electric energy while the carbon fuels may run out on all of us in just a few decades, during which time usable land may not be able to provide enough food for the growing population, cancer or dementia will strike almost everybody if we all live long enough. Well, we have sent humans to the moon; we have completely eradicated smallpox and almost done with polio, can technologies once again enable us to do big things if we all aim high and pull together?

The success stories of future technology companies should not be only the types of Facebook or Twitter, which are nice stories on their own values, but success stories should also include those that deal with big, material, and imminent challenges, provide tools that help people in desperate need. Examples in our biomedical field could include diagnostic kits based on genomic information that will one day be put into each household, so that everybody will be able to decide and receive the most suitable treatment when having an ailment. New businesses will merge because of the technology advancements of deep sequencing, information storage and analysis, biosensors, and stem cell-derived assays and delivery vehicles.

Technologies will continue to develop at a faster pace than most people’s imagination as long as there is a culture that encourages it and a system that allows those with the extraordinary ambition and brains to take their risks. As an example in one of our specific fields, the barriers to making induced pluripotent stem cells (iPSCs) have been dramatically lowered through several generations of method revolution only 6 years after the Nobel Prize-winning discovery was first published in 2006 because researchers believe that there will be new opportunities if reprogramming can be done more efficiently and “cleanly”. We have contributed our share of innovation in 2012 and our ambition is to provide everybody with his or her own pluripotent stem cells ready for medical use and to find a solution to most diseases with each individual’s own tissue-derived cells, in another term, point-of-care autologous treatment. It’s unproven, it’s futuristic, but it’s exciting and feasible and we will put every effort to make it happen. Theodore Roosevelt once said that “Far and away the best prize that life has to offer is the chance to work hard at work worth doing.” We are the lucky few.

Tags: , , , , , , , , ,

Saturday, December 29th, 2012 Open Forum No Comments

American CryoStem Corporation (OTCQB:CRYO), announced the launch of its newest adult stem cell and adipose tissue collection center in Bellevue, Washington

A public company doing business of preparing and providing adipose (fat) tissue and adipose derived adult stem cells, American CryoStem Corporation (OTCQB:CRYO), announced the launch of its newest adult stem cell and adipose tissue collection center in Bellevue, Washington. Dr. Fredric Stern will officially launch the new Stern Center Stem Cell Collection Service as the first to provide Adult Stem Cell and Tissue Banking services to the general public in the Seattle, Washington area.

“Having successfully worked with American CryoStem in the past we are truly excited about the official launch of these adipose tissue based services to the general public in Washington. I look forward to working with American CryoStem on educating my patients about the Regenerative Medicine benefits of “bio-banking” and the latest fat transfer cosmetic services now available at the center. I chose to affiliate my practice with American Cryostem because of their thorough scientific approach to stem cell banking and strict adherence to aseptic technique and FDA guidelines,” said Dr. Fredric Stern, the founder of The Stern Center and a plastic surgeon.

John S. Arnone, CEO said, according to a company news release, “We are excited to have a surgeon with Dr. Stern’s abilities and reputation associated with American CryoStem in the Seattle, WA area and look forward to a productive relationship with the entire Stern Center team. We remain committed to our “Gold Standard” clinical laboratory processing and storage reputation and strive to provide the best physician and patient services in the U.S. The newest stem cell collection center in our network represents our commitment to associate with leading physicians in the Regenerative Medicine Industry.”

Mesenchymal stem cells (MSCs) are typically the products of adipose tissue-isolated stem cells for regenerative medicine or, in this case cosmetic surgeries. The mesenchymal stem cells can also be isolated from bone marrow or embryos. They secret hormones once introduced into human bodies and help balance cytokines in the blood. It is reported that MSCs help reduce several disease symptoms and, in some countries, are used as “youth fountains” in anti-aging treatment. MSCs can be produced fairly easily, in our hands at least, from induced pluripotent stem cells (iPSCs). iPSCs, like embryonic stem cells, can be expanded indefinitely, differentiated into MSCs and all other cell types, and are being tested in various cell therapies including those that are mediated through the MSC stage.

Tags: , , , ,

Wednesday, December 12th, 2012 iPSCs and other stem cells, Open Forum No Comments