Kobolak Julianna, Bodo Szilard, Rungsiwiwut Ruttachuk, Meng Qinggang, Adorjan Marta, Virutamasen Pramuan, Techakumphu Mongkol, Dinnyes Andras
Micromanipulation and Genetic Reprogramming Group, Agricultural Biotechnology Center, Gödöllo, Hungary.
Cell Reprogram. 2010 Feb;12(1):105-13. doi: 10.1089/cell.2009.0040.
Pluripotent stem cells would have great potential in cell therapies and drug development when genetically matched with the patient; thus, histocompatible cells could be used in transplantation therapy or as a source of patient-specific cells for drug testing. Pluripotent embryonic stem cells (ESCs)-generated via somatic cell nuclear transfer (SCNT) or parthenogenesis (pESC)-are potential sources of histocompatible cells and tissues for transplantation. Earlier studies used the piezoelectric microinjection (PEM) technique for nuclear transfer (NT) in mouse. No specific studies examined zona-free (ZF) NT as an alternative NT method to generate genetically matched ESCs of a nuclear donor. In this study, we compared the efficiency of nuclear transfer-derived ESC (ntESC) line establishment from ZF-NT, ZF-parthenogenetic (PGA), and ZF-fertilized embryos with that of the PEM-NT method. Different nuclei donor cells [cumulus, ESC, and mouse embryonic fibroblast (MEF)] were used and the efficiency of ntESC derivation was investigated, along with their in vitro characterization. The ZF-NT method's efficiency was higher than that of the PEM-NT using cumulus cells. When ESCs and cumulus cells were used as nuclear donor cells, they resulted in significantly higher ZF-NT-derived ntESC line establishment rates compared to MEF cells. In conclusion, the nuclear donor cell type significantly affected the efficiency of ntESC line establishment, and the ZF-NT method was efficient to establish pluripotent ntESC lines.
多能干细胞在与患者基因匹配时，在细胞治疗和药物开发方面具有巨大潜力；因此，组织相容性细胞可用于移植治疗或作为药物测试中患者特异性细胞的来源。通过体细胞核移植（SCNT）或孤雌生殖（pESC）产生的多能胚胎干细胞（ESC）是用于移植的组织相容性细胞和组织的潜在来源。早期研究在小鼠中使用压电显微注射（PEM）技术进行核移植（NT）。没有具体研究将无透明带（ZF）核移植作为一种替代核移植方法来产生核供体的基因匹配ESC。在本研究中，我们比较了从ZF核移植、ZF孤雌生殖（PGA）和ZF受精胚胎建立核移植来源的ESC（ntESC）系的效率与PEM核移植方法的效率。使用了不同的核供体细胞[卵丘细胞、ESC和小鼠胚胎成纤维细胞（MEF）]，研究了ntESC衍生的效率及其体外特性。ZF核移植方法的效率高于使用卵丘细胞的PEM核移植。当ESC和卵丘细胞用作核供体细胞时，与MEF细胞相比，它们导致ZF核移植衍生的ntESC系建立率显著更高。总之，核供体细胞类型显著影响ntESC系建立的效率，并且ZF核移植方法对于建立多能ntESC系是有效的。
