Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells

摘要

The transfer of somatic cell nuclei into oocytes can give rise to pluripotent stem cells that are consistently equivalent to embryonic stem cells, holding promise for autologous cell replacement therapy. Although methods to induce pluripotent stem cells from somatic cells by transcription factors are widely used in basic research, numerous differences between induced pluripotent stem cells and embryonic stem cells have been reported, 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.%以前的研究表明，多能干细胞系可以通过"体细胞核转移"（SCNT,-个成熟细胞的核被转移到一个未受精的、不成熟的卵细胞中）来从人类胚胎和新生儿成纤维细胞（皮肤细胞）获得。现在，Dieter Egli及同事报告了胚胎干细胞系以SCNT方式从成年体细胞(包括一个1-型糖尿病供者的皮肤细胞）的成功获得。他们的研究还系统性地考察了在其获取干细胞系的过程中影响效率和发育潜力的参数，这些参数对于潜在治疗应用的程序改进也许有重要意义。