Culture medium refinement by dialysis for the expansion of human induced pluripotent stem cells in suspension culture
| Autor: | Eiji Nagamori, Suman Chandra Nath, Masanobu Horie, Masahiro Kino-oka |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Basic fibroblast growth factor Induced Pluripotent Stem Cells Cell Culture Techniques Bioengineering 01 natural sciences 03 medical and health sciences chemistry.chemical_compound Adherent Culture 010608 biotechnology Humans Secretion Autocrine signalling Growth medium biology business.industry General Medicine Transforming growth factor beta Biotechnology Lactic acid Cell biology Culture Media 030104 developmental biology chemistry biology.protein Dialysis (biochemistry) business Dialysis |
| Zdroj: | Bioprocess and biosystems engineering. 40(1) |
| ISSN: | 1615-7605 |
| Popis: | Human induced pluripotent stem cells (hiPSCs) secrete essential autocrine factors that are removed along with toxic metabolites when the growth medium is exchanged daily. In this study, after determining the minimum inhibitory level of lactic acid for hiPSCs, a medium refining system was constructed by which toxic metabolites were removed from used culture medium and autocrine factors as well as other growth factors were recycled. Specifically, about 87 % of the basic fibroblast growth factor and 80 % of transforming growth factor beta 1 were retained in the refined medium after dialysis. The refined medium efficiently potentiated the proliferation of hiPS cells in adherent culture. When the refining system was used to refresh medium in suspension culture, a final cell density of (1.1 ± 0.1) × 106 cells mL−1 was obtained, with 99.5 ± 0.2 % OCT 3/4 and 78.3 ± 1.1 % TRA-1-60 expression, on day 4 of culture. These levels of expression were similar to those observed in the conventional suspension culture. With this method, culture medium refinement by dialysis was established to remove toxic metabolites, recycle autocrine factors as well as other growth factors, and reduce the use of macromolecules for the expansion of hiPSCs in suspension culture. |
| Databáze: | OpenAIRE |
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