Analysis of the distinct functions of growth factors and tissue culture substrates necessary for the long-term self-renewal of human embryonic stem cell lines
| Autor: | Patricia Murray, Nicola Bates, Fiona Small, Melissa A. Baxter, Susan J. Kimber, Maria V. Camarasa, David Edgar |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Integrin
Self renewal Fibroblast growth factor Culture Media Serum-Free Cell Line 03 medical and health sciences Tissue culture 0302 clinical medicine Humans Nerve Growth Factors Embryonic Stem Cells reproductive and urinary physiology Cell Proliferation 030304 developmental biology Medicine(all) 0303 health sciences biology Integrin beta1 Cell Differentiation General Medicine Cell Biology Embryonic stem cell Activins Fibronectins Cell biology Fibronectin Cell culture embryonic structures biology.protein Fibroblast Growth Factor 2 biological phenomena cell phenomena and immunity 030217 neurology & neurosurgery Neurotrophin Developmental Biology |
| Zdroj: | Stem Cell Research. (1):28-38 |
| ISSN: | 1873-5061 |
| DOI: | 10.1016/j.scr.2009.03.002 |
| Popis: | The role of individual supplements necessary for the self-renewal of human embryonic stem (hES) cells is poorly characterized, and furthermore we have found that previously reported feeder cell- and serum-free culture systems used for individual hES cell lines are unable to maintain HUES7 cells for more than one passage. We have therefore derived a feeder/serum-free culture system that can support the long-term (at least 10 passages) self-renewal of several euploid hES cell lines including MAN1, HUES7, and HUES1 with minimal spontaneous differentiation and without the need for manual propagation. This system contains fibroblast growth factor 2, activin A, neurotrophin 4, and the N2, B27 supplements together with a human fibronectin substrate. We demonstrate that these components exert distinct functions: both FGF2 and activin A were necessary to prevent differentiation of hES cells while NT4 promoted cell survival, FGF2 could not be substituted by IGFII, and the fibronectin substrate supported a rapid rate of hES culture expansion. Inhibition studies showed that β1 integrin-dependent attachment of hES cells to fibronectin was at least partially via the α5 subunit but independent of integrin αV. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|