Derivation and characterization of human embryonic stem cell lines from poor quality embryos.
Poor quality embryos discarded from in vitro fertilization (IVF) laboratories are good sources for deriving human embryonic stem cell (hESC) lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses (ICMs) could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods (P>0.05). As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.
Guangzhou Key Laboratory of Reproductive and Genetics; Institute of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, China.
Weiqiang
W
Liu
Yifei
Y
Yin
Xiaolin
X
Long
Yumei
Y
Luo
Yonghua
Y
Jiang
Wenhong
W
Zhang
Hongzi
H
Du
Shaoying
S
Li
Yuhong
Y
Zheng
Qing
Q
Li
Xinjie
X
Chen
Baoping
B
Liao
Guohong
G
Xiao
Weihua
W
Wang
Xiaofang
X
Sun
D053624:Blastocyst Inner Cell Mass / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008297:Male