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Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes

Cell Research 2003年第4期13卷 251-263页

作者： YING CHEN, ZHI Xu HE, AILIAN LIU, KAI WANG, WEN WEI MAO, JIAN XIN CHU, YONG LU, ZHENG Fu FANG, YING TANG SHI, QING ZHANG YANG, DA YUAN CHEN, MIN RANG WANG, JIN SONG LI, SHAO LIANG HUANG, XIANG YIN KONG, YAO ZHOU SHI, Zm QIANG WANG, JIA HuI XIA, ZHI GAO LONG, ZHI GANG XUE, WEN XIANG DING, HuI ZHEN SHENG Center for Developmental Biology, Shanghai Second Medical University, Shanghai 200092, China Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 1665 Kong Jiang Road, Shanghai 200092, China Center for Stem Cell Research, the Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510120,China State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 19 Zhong-guan Cun Road, Haidian District, Beijing 100080, China Health Science Center, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China National Laboratory of Medical Genetics, Central South University, 88 Xiangya Road, Changsha 410078, China Xinhua Hospital, Shanghai Second Medical University, 1665 Kong Jiang Road, Shanghai 200092, China 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 3Center for Stem Cell Research the Second Affiliated Hospital Sun Yat-Sen University Guangzhou 510120 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 2Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 1665 Kong Jiang Road Shanghai 200092 China. 4State Key Laboratory of Reproductive Biology Institute of Zoology Chinese Academy of Sciences 19 Zhong-guan Cun Road Haidian District Beijing 10008

To solve the problem of immune incompatibility, nuclear transplantation has been envisaged as a means to produce cells or tissues for human autologous transplantation. Here we have derived embryonic stem cells by the transfer of human somatic nuclei into rabbit oocytes. The number of blastocysts that developed from the fused nuclear transfer was comparable among nuclear donors at ages of 5, 42, 52 and 60 years, and nuclear transfer (NT) embryonic stem cells (ntES cells) were subsequently derived from each of the four age groups. These results suggest that human somatic nuclei can form ntES cells independent of the age of the donor. The derived ntES cells are human based on karyotype, isogenicity, in situ hybridization, PCR and immunocytochemistry with probes that distinguish between the various species. The ntES cells maintain the capability of sustained growth in an undifferentiated state, and form embryoid bodies, which, on further induction, give rise to cell types such as neuron and muscle, as well

关键词： nuclear transfer (NT) somatic cell nuclear transfer (SCNT) embryonic stem cells (ES cell) therapeutic cloning rabbit oocyte.

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Experimental cloning of embryos through human-rabbit inter-species nuclear transfer

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Frontiers in Biology 2007年第1期2卷 80-84页

作者： JI Jingjuan GUO Tonghang TONG Xianhong LUO Lihua ZHOU Guixiang FU Yingyun LIU Yusheng Center for Reproductive Medicine Anhui Medical University Affiliated Provincial HospitalHefei 230001China

therapeutic cloning,which is based on human somatic cell nuclear transfer,is one of our major research *** inter-species nuclear transfer has been introduced to construct human somatic cell cloned embryos,the effects of type,passage,and preparation method of donor cells on embryo development remain *** our experiment,cloned embryos were reconstructed with different passage and preparation methods of ossocartilaginous cell,skin fibroblast,and cumulus *** cumulus cell embryos showed significantly higher development rates than the other two(P<0.05).The development rate of embryos reconstruct-ed with skin fibroblasts of different passage number and somatic cells of different chilling durations showed no significant ***,fluorescence in situ hybridization(FISH)was conducted to detect nuclear derivation of the *** result showed that the nuclei of the inter-species cloned embryo cells came from *** conclude that(1)cloned embryos can be constructed through human-rabbit interspecies nuclear transfer;(2)different kinds of somatic cells result in different efficiency of nuclear transfer,while in vitro passage of the donor does not influence embryo development;(3)refrigeration is a convenient and efficient donor cell preparation ***,it is feasible to detect DNA genotype through FISH.

关键词： therapeutic cloning nuclear transfer somatic cell nuclear transfer inter-species cloned embryo

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Establishment of customized mouse stem cell lines by sequential nuclear transfer

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Cell Research 2007年第1期17卷 80-87页

作者： Alice Jouneau FRE 2857 Biologie du Développement et Reproduction UMR 1198 ENVA CNRS F-78350 INRA Chinese Academv of Sciences Jouy en Josas Beijing 100080 France State Key Laboratory of Reproductive BiologyJoint laboratory of biology of embryonic cells of mammalians(LABIOCEM) Institute of Zoology China

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

关键词： nuclear transfer therapeutic cloning embryonic stem cells

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