Methods for expansion of human embryonic stem cells.
The manipulation of human embryonic stem cells (hESCs) requires refined skills. Here we introduce both mechanical and enzymatic transfer methods for hESCs depending on experimental purpose. We use the mechanical transfer method for maintenance of hESC lines. Although the method is laborious and time-consuming, the technique permits efficient transfer of undifferentiated hESCs and results in similar clump sizes. We implement the enzymatic transfer method when we need the bulk production of cells for various experiments. The enzyme-treated expansion rapidly produces greater amounts of hESCs within a limited time frame. However, the cell clumps vary in size, and there is a probability that both the differentiated and undifferentiated cells will be transferred. In cases in which there are differentiated colonies, the combination of two methods allows mass production of hESCs by excluding differentiated colonies from passage by manual selection before enzyme treatment.
Oh
Sun Kyung
SK
Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-744, Korea.
Kim
Hee Sun
HS
Park
Yong Bin
YB
Seol
Hye Won
HW
Kim
Yoon Young
YY
Cho
Myung Soo
MS
Ku
Seung Yup
SY
Choi
Young Min
YM
Kim
Dong-Wook
DW
Moon
Shin Yong
SY
D018929:Cell Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology*; D006801:Humans; D013234:Stem Cells / Q000166:cytology*