Isolation, cryopreservation and culture of human amnion epithelial cells for clinical applications

• Published in: Journal of visualized experiments 2014-12 (94)
• Main Authors: Murphy, Sean V, Kidyoor, Amritha, Reid, Tanya, Atala, Anthony, Wallace, Euan M, Lim, Rebecca
• Format: Article
• Language: English
• Subjects: Amnion - cytology; Cell Differentiation; Cell Separation - methods; Cell- and Tissue-Based Therapy; Cryopreservation; Epithelial Cells - cytology; Humans; Medicine; Regenerative Medicine
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/52085

Human amnion epithelial cells (hAECs) derived from term or pre-term amnion membranes have attracted attention from researchers and clinicians as a potential source of cells for regenerative medicine. The reason for this interest is evidence that these cells have highly multipotent differentiation ability, low immunogenicity, and anti-inflammatory functions. These properties have prompted researchers to investigate the potential of hAECs to be used to treat a variety of diseases and disorders in pre-clinical animal studies with much success. hAECs have found widespread application for the treatment of a range of diseases and disorders. Potential clinical applications of hAECs include the treatment of stroke, multiple sclerosis, liver disease, diabetes and chronic and acute lung diseases. Progressing from pre-clinical animal studies into clinical trials requires a higher standard of quality control and safety for cell therapy products. For safety and quality control considerations, it is preferred that cell isolation protocols use animal product-free reagents. We have developed protocols to allow researchers to isolate, cryopreserve and culture hAECs using animal product-free reagents. The advantage of this method is that these cells can be isolated, characterized, cryopreserved and cultured without the risk of delivering potentially harmful animal pathogens to humans, while maintaining suitable cell yields, viabilities and growth potential. For researchers moving from pre-clinical animal studies to clinical trials, these methodologies will greatly accelerate regulatory approval, decrease risks and improve the quality of their therapeutic cell population.