Richards Mark, Tan Shawna, Fong Chui-Yee, Biswas Arjit, Chan Woon-Khiong, Bongso Ariff
Department of Obstetrics and Gynecology, National University of Singapore, Kent Ridge, Singapore.
Stem Cells. 2003;21(5):546-56. doi: 10.1634/stemcells.21-5-546.
Human embryonic stem (hES) cells are typically derived and serially propagated on inactivated murine embryonic fibroblast (MEF) feeders. The use of MEFs and other components of animal origin in the culture media for hES cell support substantially elevates the risk of contaminating these cell lines with infectious agents of animal origin thereby severely limiting their potential for clinical application. We have previously shown that it is possible to derive and establish new hES cell lines in a xeno-free culture system using human fetal muscle fibroblast feeders. In this report, we have comparatively evaluated a panel of 11 different human adult, fetal, and neonatal feeders for hES cell support and have ranked them as supportive and non-supportive. We report that two adult skin fibroblast cell lines established in-house from abdominal skin biopsies supported prolonged undifferentiated hES cell growth for over 30 weekly passages in culture. Furthermore, hES cell lines cultured on adult skin fibroblast feeders retain hES cell morphology and remain pluripotent. Also, differences in feeder support exist between human cell types and sources. The use of human adult skin feeders is convenient for hES cell support given the ease of obtaining skin biopsies.
人胚胎干细胞（hES细胞）通常在灭活的小鼠胚胎成纤维细胞（MEF）饲养层上获得并进行传代培养。在用于支持hES细胞培养的培养基中使用MEF及其他动物源成分，会大幅增加这些细胞系被动物源感染因子污染的风险，从而严重限制其临床应用潜力。我们之前已表明，使用人胎儿肌肉成纤维细胞饲养层，在无血清培养系统中能够获得并建立新的hES细胞系。在本报告中，我们比较评估了11种不同的人成年、胎儿和新生儿饲养层对hES细胞的支持作用，并将它们分为支持性和非支持性两类。我们报告称，从腹部皮肤活检样本中自行建立的两种成人皮肤成纤维细胞系，支持hES细胞在培养中持续30周以上的未分化生长。此外，在成人皮肤成纤维细胞饲养层上培养的hES细胞系保持hES细胞形态并维持多能性。而且，人细胞类型和来源之间在饲养层支持作用方面存在差异。鉴于获取皮肤活检样本较为容易，使用成人皮肤饲养层来支持hES细胞培养很方便。
