Yamato et al., "Human adipose tissue-derived mesenchymal stem cells as a novel feeder layer
for epithelial cells," Journal of Tissue Engineering and Regenerative Medicine, vol.
del Rio, "Feeder layer
cell actions and applications," Tissue Engineering.
Suzuki et al., "Human placenta feeder layers
support undifferentiated growth of primate embryonic stem cells," Stem Cells, vol.
Generating safer and more efficient cells using more appropriate culture media and feeder layers
provides more adequate cells for cell transplantation therapies.
Vigorously pipetted up and down to obtain single cells from plucked hair, and then centrifuged at 200xg for 5 them for specific period of time in serum based medium without feeder layer
We now know that standardization of the method with human originated feeder layer
cells and use of animal-free medium are necessary to use these cells in therapeutic applications.
Previous studies have shown that human limbal epithelial progenitor cells can be expanded and maintained on intact, but not denuded HAM or plastic in the absence of 3T3 murine fibroblast feeder layers
. Intrestingly He et al (22) has demonstrated the role of PI3K/Akt signaling pathways on the cells expanded over the intact membrane.
Presently, appropriate cytokines and feeder layers
are widely applied to keep cESCs undifferentiated.
The results showed that some keratinocytes in monolayer cultures highly expressed C4.4A at cell-cell borders (Figures 1(a)-1(b), arrows), while the murine 3T3 fibroblasts, used as a feeder layer
in the culture, were devoid of reactivity (Figure 1(a), arrowhead).
After electroporation, mESCs were allowed to spread on pre-coated gelatin-plastic flasks containing mitomycin C-inactivated feeder layer
of primary cultures of mouse embryonic fibroblasts (MEF) in ESC medium containing G418 (Sigma, USA).
(connexin 43 and keratin 3 / keratin 12) on the limbal epithelial cells cultured over the denuded,AM with and without the 3T3 murine fibroblast cells as feeder layer
Briefly, mitomycin c- (MMC- ; Sigma-Aldrich) treated (20 [micro]g/ml, for 3 h) mouse bone marrow stromal cells (M210B4; Korean Cell Line Bank, Seoul, South Korea) were seeded at 3 x [10.sup.5] cells/well in a 35 mm culture dish as a feeder layer
the day before the assay.