TISSUE CULTURE, QUALITY ASSURANCE WITH GMP, ESSENTIAL FOR HUMAN CELL THERAPY
For successful human cell therapy, quality cells must be ensured by the laboratory and conform to GMP, sterility and quality assurance regards tissue markers etc. Normally this cell expansion is only done in an academic unit where all the expertise, senior control, audit and ongoing studies in cell biology are present. Special ethical approval is mandatory, and the principal investigator must be a specialist clinician at doctorate level in cell biology, because of the need for patient care and oversight of the culture process, and audit. The process is basic:
- Tissue biopsy or tissue harvest. This is a special task for a specialist to reduce morbidity
- Tissue transport to the lab in special mediums
- Tissue processing, and plating in special tissue culture plates or flasks. Special mediums are used in which to grow the cells
- Cells are proliferated and expanded in a GMP tissue culture laboratory that maintains sterility
- Sufficient numbers of cells are expanded ex vivo, cultivated, careful observation daily
- Cell cultivation takes 4-12 weeks before they can be used. Some cells from donor biopsies don’t grow because of various reasons. Medical -qualified cell biology specialists are needed to audit this problem
- Cells are tested for viability and tissue markers
- Once sufficient cells are expanded, cells are released from the tissue culture process, and the cells are placed in a special cell culture transplant medium
- The cells are delivered to the specialist who performs the cell transplantation
- Provided recipient tissue enhances cell retention the new cell will grow and proliferate and rejuvenate or resurface the defective or aged organ (say the skin can be resurfaced with new cells to ameliorate wrinkles and the aging process: this is referred to as the “Isolagen Process”. This is not stem-cell therapy.
- Cell therapy is a complex matter, labour intensive and cost ineffective at the moment compared to other treatments. Costs often exceed R25,000 per treatment
- Cells and tissues may not cross borders of South Africa, which is controlled by The Human Tissue Act within the National Health Act of 2003.That means tissues from other countries may not be processed in South Africa or sent abroad.
- Special ethical approval for protocols, and long periods of cell storage are forbidden as well as attempting to clone humans. Numerous cell biotechnology patents are in place and this restricts cell proliferation on a random basis
- Cells that are currently in stages of investigation include: fibroblasts (old burns and scars), myoblasts (damaged myocardium), chondrocytes (joint degeneration) and keratinocytes (burns, vitelligo). Each cell line has its own phenotype and the culture procedure needs to be overseen by an experienced cell-based and trained medically qualified health care professional with experience in cell transplantation immunology and potential side-effects in cell therapy. Just transplanting cells obtained from an obscure cell laboratory, by non-specialists may harm the public, as has been seen in the past. The most popular process has been fibroblast proliferation for “THE ISOLAGEN PROCESS”. This process was further pioneered in Cape Town by Du Toit and co-workers during 2005-2008. Chondrocytes have been moderately successful for joint resurfacing.
- Cell culture mediums may consist of any one of the following: Collagen and Matrigel® bases, Eagles®, Ham’s and DMEM solutions either as serum free or enrichment with autologous serum, foetal calf or bovine serum, basic fibroblast growth factor. Fibronectin coated plates, as adhesion molecules can also be used. Platelet-rich plasma (PRP, 5, 10, 20% in serum free mediums as a patented process), from the same tissue donor, has been in use for more than 3 years. Others include Oligene® medium for fibroblasts, keratinocytes or chondrocytes. Cell growth may be carefully controlled by the use of red light. Static or dynamic culture is possible in 3-D dimension and may produce a better quality cell than generated in static, monolayer culture.
- The most advanced biotechnology facilty currently operaing in the field of cell technology and therapy and supported by evidence–based medicine, peer-review environment, audit registries, standard-orders-and-procedures, and academic publications, has been directed by a medical specialist Donald du Toit, in Cape Town, the only BIOMED rated expert in this field in South Africa as of 2007.
REFERENCES relevant to biotechnology by BIOMED EXPERTS:
- Du Toit DF et al. Biotechnological anti-aging cell-therapy treatment of facial wrinkles with cultured human fibroblasts. The Specialist Forum: 5:38-46, 2005
- Boss WK et al. Autologous cultured fibroblasts as cellular therapy in plastic surgery. Clinics in plastic surgery 27:613-626, 2000
- Boss et al. Autologous cultured fibroblasts: A protein repair system. Ann Plast Surg 44:536-542, 2000.
WEBSITE UPDATED AND POSTED 20 OCTOBER 2008.
CONSULTANT BIOMED EXPERT FORMULATED CELL-BASED TISSUE CULTURE TECHNOLOGIES
CELL TISSUE GALLERY REFLECTING MORPHOLOGY OF CULTURED CELLS