title: Establishment of customized mouse stem cell lines by sequential nuclear transfer.
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.
authors.affiliation: State Key Laboratory of Reproductive Biology, Chinese Academy of Sciences, Beijing 100080, China.
authors.forename: Chunli
authors.initials: C
authors.lastname: Zhao
authors.forename: Ruqiang
authors.initials: R
authors.lastname: Yao
authors.forename: Jie
authors.initials: J
authors.lastname: Hao
authors.forename: Chenhui
authors.initials: C
authors.lastname: Ding
authors.forename: Yong
authors.initials: Y
authors.lastname: Fan
authors.forename: Xiangpeng
authors.initials: X
authors.lastname: Dai
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Tang
authors.initials: T
authors.lastname: Hai
authors.forename: Zichuan
authors.initials: Z
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Yingying
authors.initials: Y
authors.lastname: Wang
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Hou
authors.forename: Weizhi
authors.initials: W
authors.lastname: Ji
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.forename: Alice
authors.initials: A
authors.lastname: Jouneau
authors.forename: Fanyi
authors.initials: F
authors.lastname: Zeng
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
mesh_terms: D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology*; D019976:Cloning, Organism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D053652:Nuclear Transfer Techniques