Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells
Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells
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人类卵母细胞将1型糖尿病患者的成人体细胞核重编程为二倍体多能干细胞。
DOI：
10.1038/nature13287
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发表时间：
2014-06-26
期刊：
影响因子：
64.8
通讯作者：
Egli, Dieter
中科院分区：
文献类型：
--
作者：
Yamada, Mitsutoshi;Johannesson, Bjarki;Egli, Dieter
将体细胞核转移到卵母细胞中可以产生与胚胎干细胞一致的多能干细胞(1-3)，这为自体细胞替代疗法带来了希望(4,5)。尽管通过转录因子从体细胞诱导多能干细胞的方法(6) 广泛应用于基础研究中，但诱导多能干细胞和胚胎干细胞之间存在许多差异已被报道(7-11)，这可能会影响其临床应用。由于源自患病人类受试者成体细胞的二倍体胚胎干细胞系的治疗潜力，我们系统地研究了影响囊胚发育和干细胞衍生效率的参数。在这里，我们展示了卵母细胞激活方案的改进，包括使用激酶和翻译抑制剂，以及在组蛋白脱乙酰酶抑制剂存在下进行细胞培养，可促进囊胚阶段的发育。卵母细胞捐赠者之间的发育效率存在差异，并且与卵母细胞成熟所需的激素刺激天数成反比，而促性腺激素的每日剂量或取出的中期 II 卵母细胞总数并不影响发育结果。由于使用浓缩的仙台病毒进行细胞融合会导致细胞内钙浓度增加，导致卵母细胞过早活化，因此我们在无钙培养基中使用稀释的仙台病毒。使用这种改良的核转移方案，我们从新生儿的体细胞中衍生出二倍体多能干细胞系，并且首次从患有 1 型糖尿病的成年女性的体细胞中衍生出二倍体多能干细胞系。
The transfer of somatic cell nuclei into oocytes can give rise to pluripotent stem cells that are consistently equivalent to embryonic stem cells(1-3), holding promise for autologous cell replacement therapy(4,5). Although methods to induce pluripotent stem cells from somatic cells by transcription factors(6) are widely used in basic research, numerous differences between induced pluripotent stem cells and embryonic stem cells have been reported(7-11), potentially affecting their clinical use. Because of the therapeutic potential of diploid embryonic stem-cell lines derived from adult cells of diseased human subjects, we have systematically investigated the parameters affecting efficiency of blastocyst development and stem-cell derivation. Here we show that improvements to the oocyte activation protocol, including the use of both kinase and translation inhibitors, and cell culture in the presence of histone deacetylase inhibitors, promote development to the blastocyst stage. Developmental efficiency varied between oocyte donors, and was inversely related to the number of days of hormonal stimulation required for oocyte maturation, whereas the daily dose of gonadotropin or the total number of metaphase II oocytes retrieved did not affect developmental outcome. Because the use of concentrated Sendai virus for cell fusion induced an increase in intracellular calcium concentration, causing premature oocyte activation, we used diluted Sendai virus in calcium-free medium. Using this modified nuclear transfer protocol, we derived diploid pluripotent stem-cell lines from somatic cells of a newborn and, for the first time, an adult, a female with type 1 diabetes.