A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells

Abstract

In order to apply human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) to regenerative medicine, the cells should be produced under restricted conditions conforming to GMP guidelines. Since the conventional culture system has some issues that need to be addressed to achieve this goal, we developed a novel culture system. We found that recombinant laminin-511 E8 fragments are useful matrices for maintaining hESCs and hiPSCs when used in combination with a completely xeno-free (Xf) medium, StemFit™. Using this system, hESCs and hiPSCs can be easily and stably passaged by dissociating the cells into single cells for long periods, without any karyotype abnormalities. Human iPSCs could be generated under feeder-free (Ff) and Xf culture systems from human primary fibroblasts and blood cells, and they possessed differentiation abilities. These results indicate that hiPSCs can be generated and maintained under this novel Ff and Xf culture system.

Animal‐cell culture media: History, characteristics, and current issues

“…These cells initially were cultivated on a layer of feeder cells in a serum‐ or KSR‐supplemented medium,104 but in anticipation of clinical applications, the efforts shifted to the development of culture conditions that are feeder‐free and xeno‐free (Table 5). 105, 106, 107, 108, 109, 110, 111 Among these, the E8 medium that was developed by Guokai Chen et al. has become popular.…”

Применение Плюрипотентных Стволовых Клеток В Медицине

“…The latter is an important issue, because a contact with components of animal origin may induce immunogenicity of hPSCs and their differentiated progeny [36]. To date, methods of derivation, culturing and freezing/thawing under xeno-free (devoid of components of animal origin) and chemically defined conditions have been reported for hESCs [37][38][39][40][41][42] and hiPSCs [43,44]. The methods comply with the most stringent rules and it will be easy to receive an approval of regulatory agencies for treatments based on them.…”

Cell therapy for Parkinson’s disease

“…[30][31][32] now, researchers can prepare iPs cells from peripheral blood cells, which is less invasive than skin biopsy, by gene transfection without genome insertion or the use of oncogenes like c-Myc, and without the use of feeder cells. 32,33) Consequently, iPs cells compatible with clinical application are now available.…”