The expansion of human ES and iPS cells on porous membranes and proliferating human adipose-derived feeder cells.
For clinical application of human embryonic stem cells (hESCs), it is critical to develop hESC culture techniques that completely exclude the use of animal feeder cells, mitotic inhibition, and enzyme treatments used in conventional hESC culture systems. Toward this goal, we attempted to maintain hESCs and induced pluripotent stem (iPS) cells on porous membranes (PMs) with proliferative human adipose-derived stromal cells (ASCs) seeded on the bottom surface of inverted PMs. This culture condition will ensure that the two cell types are separate from each other, yet retain the ability to interact through the pores of the membrane. We found that hESCs and iPS cells can be maintained stably and mechanically transferred without the need for enzyme treatment. In addition, the pluripotency of hESCs and iPS cells was stably maintained, as evidenced by immunostaining of Oct4, SSEA3/4 and TRA-1-60 as well as RT-PCR analyses of Nanog, Oct4 and Sox2 expression. Furthermore, hESCs cultured on PMs showed a normal karyotype and in vivo teratoma formation containing all three germ layers.
Hwang
Seung-Taeh
ST
Department of Biomedical Science, CHA Stem Cell Institute, CHA University, Seoul, Republic of Korea.
Kang
Sun-Woong
SW
Lee
Suk-Jun
SJ
Lee
Tae-Hee
TH
Suh
Wonhee
W
Shim
Sung Han
SH
Lee
Dong-Ryul
DR
Taite
Lakeshia J
LJ
Kim
Kwang-Soo
KS
Lee
Soo-Hong
SH
D000273:Adipose Tissue / Q000166:cytology* / Q000648:ultrastructure; D049109:Cell Proliferation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D007621:Karyotyping; D008567:Membranes, Artificial; D016062:Porosity; D017154:Stromal Cells / Q000166:cytology; D013499:Surface Properties; D013724:Teratoma / Q000473:pathology
D008567:Membranes, Artificial