Illmensee Karl, Levanduski Mike, Zavos Panayiotis M
Reprogen, Limassol, Cyprus.
Fertil Steril. 2006 Apr;85 Suppl 1:1248-60. doi: 10.1016/j.fertnstert.2005.10.026.
To examine the embryonic preimplantation potential of human adult somatic cells by creating interspecies embryos via somatic cell nuclear transfer (SCNT) using bovine oocytes.
Prospective study.
Research facility of Reprogen.
PATIENT(S): Infertile couples.
INTERVENTION(S): Enucleated bovine oocytes were fused via SCNT with either human granulosa (HG) or fibroblast (HF) cells and cultured in vitro. Polymerase chain reaction (PCR) and DNA analysis were performed on the interspecies embryos. Parthenogenetically activated embryos served as controls.
MAIN OUTCOME MEASURE(S): Embryonic preimplantation development after interspecies SCNT.
RESULT(S): From enucleated bovine oocytes fused with HG cells (n = 48) and HF cells (n = 75), 15 HG- and 22 HF-derived embryos developed, some of which progressed to blastocysts (31.3% vs. 29.3%, respectively). The PCR and DNA analysis showed that the interspecies embryos contained human genomic DNA specific for the individual DNA profile of the HG or HF donor cells used for SCNT. In addition, both bovine- and human-specific mitochondrial DNA was detectable in the interspecies embryos up to the blastocyst stage. Parthenogenetic development was 46.8% and 64.9% for the HG and HF series, respectively. The SCNT efficiency index, defined as the ratio of SCNT and parthenogenetic success rate, was 66.8% for HG cells and 45.5% for HF cells.
CONCLUSION(S): This interspecies bioassay can be utilized to determine and assess the embryonic preimplantation potential of different types of human adult somatic cells.
通过使用牛卵母细胞进行体细胞核移植(SCNT)来创建种间胚胎,以研究人类成体细胞的胚胎植入前潜能。
前瞻性研究。
Reprogen研究机构。
不孕夫妇。
去核牛卵母细胞通过SCNT与人类颗粒细胞(HG)或成纤维细胞(HF)融合,并在体外培养。对种间胚胎进行聚合酶链反应(PCR)和DNA分析。孤雌激活胚胎作为对照。
种间SCNT后的胚胎植入前发育情况。
与HG细胞(n = 48)和HF细胞(n = 75)融合的去核牛卵母细胞分别发育出15个HG来源和22个HF来源的胚胎,其中一些胚胎发育至囊胚阶段(分别为31.3%和29.3%)。PCR和DNA分析表明,种间胚胎含有与用于SCNT的HG或HF供体细胞个体DNA谱特异性对应的人类基因组DNA。此外,在发育至囊胚阶段的种间胚胎中均可检测到牛和人类特异性线粒体DNA。HG和HF系列的孤雌发育率分别为46.8%和64.9%。SCNT效率指数(定义为SCNT成功率与孤雌成功率之比),HG细胞为66.8%,HF细胞为45.5%。
这种种间生物测定法可用于确定和评估不同类型人类成体细胞的胚胎植入前潜能。