An article published this year in “EXPERIMENTAL AND THERAPEUTIC
MEDICINE” using our CD105‑FITC, by our customers from Instituto de Biofísica Carlos Chagas Filho,
Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brasil, in the study of
how Hair follicle-derived mesenchymal cells support undifferentiated growth of
embryonic stem cells. Congrats and Thanks.
Summary:
The aim of the present study was to
investigate whether feeder layers composed of human hair follicle-derived
mesenchymal stem cells (hHFDCs) are able to support human embryonic stem cells
(hESCs). hHFDCs and mouse embryonic fibroblasts (MEFs) were isolated and
cultured in Dulbecco's modified Eagle's medium (DMEM)/F‑12 and low‑glucose DMEM,
respectively. hHFDCs were passaged three times and subsequently characterized.
hHFDCs and MEFs were mitotically inactivated with mitomycin C for 3 h prior to
co‑culture
with H9‑hESCs.
hESCs were initially established on a mouse feeder layer, subsequently
transferred onto a human feeder layer and split every 5 days. Cell morphology,
expression of specific ‘undifferentiation’ markers and growth factors, and the
differentiation capacity of hESCs grown on the hHFDC feeder layer were
analyzed. hHFDCs are adherent to plastic, possess the classic mesenchymal stem
cell phenotype [they express cluster of differentiation (CD)90, CD73 and CD105]
and are able to differentiate into adipocytes, chondroblasts and osteocytes,
indicating that these cells are multipotent. Population‑doubling time
analysis revealed that hHFDCs rapidly proliferate over 34.5 h. As a feeder
layer, hHFDC behaved similarly to MEF in maintaining the morphology of hESCs.
The results of alkaline phosphatase activity, reverse transcription‑quantitative
polymerase chain reaction analysis of the expression of pluripotency
transcription factors [octamer‑binding transcription factor 4 (Oct4), Nanog
and sex determining region Y‑box 2], and immunofluorescence assays of
markers (stage‑specific
embryonic antigen‑4 and Oct4) in hESCs co‑cultured
over hHFDC, indicated that the undifferentiated state of hESCs was preserved.
No change in the level of growth factor transcripts (bone morphogenetic protein
4, fibroblast growth factor‑2, vascular
endothelial growth factor, Pigment epithelium‑derived
factor and transforming growth factor‑β1)
was detected for either feeder layer prior to or following inactivation.
Similar phenotypes of embryoid body formation, size and morphology were
observed in the hHFDC and MEF feeders. In conclusion, hHFDC maintained hESCs in
an undifferentiated state comparable to MEF in standard conditions, which may
be an important finding regarding the establishment of stem cell-based
translational applications.
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