hESC expansion and stemness are independent of connexin forty-three-mediated intercellular communication between hESCs and hASC feeder cells.
BACKGROUND
Human embryonic stem cells (hESCs) are a promising and powerful source of cells for applications in regenerative medicine, tissue engineering, cell-based therapies, and drug discovery. Many researchers have employed conventional culture techniques using feeder cells to expand hESCs in significant numbers, although feeder-free culture techniques have recently been developed. In regard to stem cell expansion, gap junctional intercellular communication (GJIC) is thought to play an important role in hESC survival and differentiation. Indeed, it has been reported that hESC-hESC communication through connexin 43 (Cx43, one of the major gap junctional proteins) is crucial for the maintenance of hESC stemness during expansion. However, the role of GJIC between hESCs and feeder cells is unclear and has not yet been reported.


RESULTS
This study therefore examined whether a direct Cx43-mediated interaction between hESCs and human adipose-derived stem cells (hASCs) influences the maintenance of hESC stemness. Over 10 passages, hESCs cultured on a layer of Cx43-downregulated hASC feeder cells showed normal morphology, proliferation (colony growth), and stemness, as assessed by alkaline phosphatase (AP), OCT4 (POU5F1-Human gene Nomenclature Database), SOX2, and NANOG expression.


CONCLUSIONS
These results demonstrate that Cx43-mediated GJIC between hESCs and hASC feeder cells is not an important factor for the conservation of hESC stemness and expansion.
Department of Biomedical Science, CHA University, Sungnam, Republic of Korea.
Jin-Su
JS
Kim
Daekee
D
Kwon
Seung-Taeh
ST
Hwang
Dong Ryul
DR
Lee
Sung Han
SH
Shim
Hee-Chun
HC
Kim
Hansoo
H
Park
Won
W
Kim
Myung-Kwan
MK
Han
Soo-Hong
SH
Lee
D000273:Adipose Tissue / Q000166:cytology*; D002450:Cell Communication; D002460:Cell Line; D049109:Cell Proliferation; D018031:Connexin 43 / Q000378:metabolism*; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism*; D061252:Feeder Cells / Q000166:cytology* / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D006801:Humans; D034741:RNA, Small Interfering / Q000378:metabolism
D018031:Connexin 43; D034741:RNA, Small Interfering