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四倍体体细胞核移植来源的人类胚胎干细胞的特性分析

Characterization of Tetraploid Somatic Cell Nuclear Transfer-Derived Human Embryonic Stem Cells.

作者信息

Shin Dong-Hyuk, Lee Jeoung-Eun, Eum Jin Hee, Chung Young Gie, Lee Hoon Taek, Lee Dong Ryul

机构信息

Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul 06135, Korea.

Dept. of Animal Biotechnology, Konkuk University, Seoul 05020, Korea.

出版信息

Dev Reprod. 2017 Dec;21(4):425-434. doi: 10.12717/DR.2017.21.4.425. Epub 2017 Dec 31.

Abstract

Polyploidy is occurred by the process of endomitosis or cell fusion and usually represent terminally differentiated stage. Their effects on the developmental process were mainly investigated in the amphibian and fishes, and only observed in some rodents as mammalian model. Recently, we have established tetraploidy somatic cell nuclear transfer-derived human embryonic stem cells (SCNT-hESCs) and examined whether it could be available as a research model for the polyploidy cells existed in the human tissues. Two tetraploid hESC lines were artificially acquired by reintroduction of remained 1st polar body during the establishment of SCNT-hESC using MII oocytes obtained from female donors and dermal fibroblasts (DFB) from a 35-year-old adult male. These tetraploid SCNT-hESC lines (CHA-NT1 and CHA-NT3) were identified by the cytogenetic genotyping (91, XXXY,-6, t[2:6] / 92,XXXY,-12,+20) and have shown of indefinite proliferation, but slow speed when compared to euploid SCNT-hESCs. Using the eight Short Tendem Repeat (STR) markers, it was confirmed that both CHA-NT1 and CHA-NT3 lines contain both nuclear and oocyte donor genotypes. These hESCs expressed pluripotency markers and their embryoid bodies (EB) also expressed markers of the three embryonic germ layers and formed teratoma after transplantation into immune deficient mice. This study showed that tetraploidy does not affect the activities of proliferation and differentiation in SCNT-hESC. Therefore, tetraploid hESC lines established after SCNT procedure could be differentiated into various types of cells and could be an useful model for the study of the polyploidy cells in the tissues.

摘要

多倍体通过核内有丝分裂或细胞融合过程产生,通常代表终末分化阶段。其对发育过程的影响主要在两栖动物和鱼类中进行了研究,在哺乳动物模型中仅在一些啮齿动物中观察到。最近,我们建立了四倍体体细胞核移植来源的人类胚胎干细胞(SCNT-hESCs),并研究了其是否可作为人类组织中存在的多倍体细胞的研究模型。通过在使用从女性供体获得的MII期卵母细胞和一名35岁成年男性的皮肤成纤维细胞(DFB)建立SCNT-hESC的过程中重新引入残留的第一极体,人工获得了两条四倍体hESC系。这些四倍体SCNT-hESC系(CHA-NT1和CHA-NT3)通过细胞遗传学基因分型(91, XXXY, -6, t[2:6] / 92, XXXY, -12, +20)进行了鉴定,显示出无限增殖能力,但与整倍体SCNT-hESCs相比速度较慢。使用八个短串联重复序列(STR)标记,证实CHA-NT1和CHA-NT3系均包含核供体和卵母细胞供体的基因型。这些hESCs表达多能性标记,其胚状体(EB)也表达三个胚胎胚层的标记,并在移植到免疫缺陷小鼠后形成畸胎瘤。本研究表明,四倍体不影响SCNT-hESC的增殖和分化活性。因此,SCNT程序后建立的四倍体hESC系可分化为各种类型的细胞,并且可能是研究组织中多倍体细胞的有用模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b233/5769136/88da88a5f1f6/dr-21-4-425-g1.jpg

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