The significance of membrane fluidity of feeder cell-derived substrates for maintenance of iPS cell stemness

The biological activity of cell-derived substrates to maintain undifferentiated murine-induced pluripotent stem (iPS) cells was correlated to membrane fluidity as a new parameter of cell culture substrates. Murine embryonic fibroblasts (MEFs) were employed as feeder cells and their membrane fluidity...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2015-06, Vol.5 (1), p.11386-11386, Article 11386
Main Authors: Zhou, Yue, Mao, Hongli, Joddar, Binata, Umeki, Nobuhisa, Sako, Yasushi, Wada, Ken-Ichi, Nishioka, Chieko, Takahashi, Eiki, Wang, Yi, Ito, Yoshihiro
Format: Article
Language:English
Subjects:
Alkaline phosphatase
Animals
Biological activity
Cell culture
Cell Culture Techniques - methods
Contamination
Embryo fibroblasts
Embryos
Epidermal growth factor
Epidermal growth factor receptors
Feeder Cells - cytology
Feeder Cells - metabolism
Fluidity
Formaldehyde
Green fluorescent protein
Growth factor receptors
Induced Pluripotent Stem Cells - cytology
Induced Pluripotent Stem Cells - metabolism
Membrane Fluidity
Mice
Mitomycin C
Pluripotency
Stem cells
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The biological activity of cell-derived substrates to maintain undifferentiated murine-induced pluripotent stem (iPS) cells was correlated to membrane fluidity as a new parameter of cell culture substrates. Murine embryonic fibroblasts (MEFs) were employed as feeder cells and their membrane fluidity was tuned by chemical fixation using formaldehyde (FA). Membrane fluidity was evaluated by real-time single-molecule observations of green fluorescent protein-labeled epidermal growth factor receptors on chemically fixed MEFs. Biological activity was monitored by colony formation of iPS cells. Treatment with a low concentration of FA sustained the membrane fluidity and biological activity, which were comparable to those of mitomycin C-treated MEFs. The biological activity was further confirmed by sustained expression of alkaline phosphatase, SSEA-1, and other pluripotency markers in iPS cells after 3-5 days of culture on FA-fixed MEFs. Chemical fixation of feeder cells has several advantages such as providing ready-to-use culture substrates without contamination by proliferating feeder cells. Therefore, our results provide an important basis for the development of chemically fixed culture substrates for pluripotent stem cell culture as an alternative to conventional treatment by mitomycin C or x-ray irradiation.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep11386