Merck
CN
  • Establishment of customized mouse stem cell lines by sequential nuclear transfer.

Establishment of customized mouse stem cell lines by sequential nuclear transfer.

Cell research (2007-01-11)
Chunli Zhao, Ruqiang Yao, Jie Hao, Chenhui Ding, Yong Fan, Xiangpeng Dai, Wei Li, Tang Hai, Zichuan Liu, Yang Yu, Yingying Wang, Xiaojun Hou, Weizhi Ji, Qi Zhou, Alice Jouneau, Fanyi Zeng, Liu Wang
ABSTRACT

Therapeutic cloning, whereby embryonic stem cells (ESCs) are derived from nuclear transfer (NT) embryos, may play a major role in the new era of regenerative medicine. In this study we established forty nuclear transfer-ESC (NT-ESC) lines that were derived from NT embryos of different donor cell types or passages. We found that NT-ESCs were capable of forming embryoid bodies. In addition, NT-ESCs expressed pluripotency stem cell markers in vitro and could differentiate into embryonic tissues in vivo. NT embryos from early passage R1 donor cells were able to form full term developed pups, whereas those from late passage R1 ES donor cells lost the potential for reprogramming that is essential for live birth. We subsequently established sequential NT-R1-ESC lines that were developed from NT blastocyst of late passage R1 ESC donors. However, these NT-R1-ESC lines, when used as nuclear transfer donors at their early passages, failed to result in live pups. This indicates that the therapeutic cloning process using sequential NT-ESCs may not rescue the developmental deficiencies that resided in previous donor generations.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Stage-Specific Embryonic Antigen-1 Antibody, clone MC-480, clone MC-480, Chemicon®, from mouse
Sigma-Aldrich
L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
5-Bromo-4-chloro-3-indolyl phosphate p-toluidine salt, tablet