Inamdar Maneesha S, Venu Parvathy, Srinivas M S, Rao Kamini, VijayRaghavan K
Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India.
Stem Cells Dev. 2009 Apr;18(3):423-33. doi: 10.1089/scd.2008.0131.
Human embryonic stem (hES) cells are a valuable tool for studying human development in addition to their potential applications in regenerative medicine and drug discovery. The role of genetic background and epigenetic influences in development as well as in response to external influences such as drugs and therapies is well recognized. The great ethnic diversity in the Indian subcontinent translates to interindividual variability in drug response and disease susceptibility. For these reasons, new hES cell lines representing Indian genetic diversity will be valuable in studies of tissue-differentiation, cellular-function and for aspects of characterization of responses to drugs. We have derived two new hES cell lines, BJNhem19 and BJNhem20 from the inner cell mass (ICM) of discarded grade III human embryos that were not suitable for in vitro fertility treatment. Human leukocyte antigen (HLA) isotype analysis shows that they are genetically distinct from existing hES cell lines. Short tandem repeat (STR) analysis shows that the two cell lines are derived from sibling embryos. These cell lines show an undifferentiated phenotype in culture for more than 65 passages, show normal karyotype and express pluripotency markers such as TRA-1-60, TRA-1-81, stage-specific embryonic antigen-4 (SSEA-4), alkaline phosphatase, DNMT3B, GABRB3, GDF3, OCT4, NANOG, SOX2, TERF1, TDGF, LEFTA, THY1, and REX1. While both cell lines can differentiate into derivatives of all three germ layers in vitro, only BJNhem20 can form teratomas when transplanted into mice. We observe an increased frequency of cardiomyocyte differentiation from BJNhem20 embryoid bodies in feeder-free cultures upon induction with DMSO. Cardiomyocytes purified from such cultures survive and show rhythmic contractions for several weeks in culture. These hES cell lines have been accepted for deposit in the U.K. Stem Cell Bank and will be a useful resource for the international stem cell community.
人类胚胎干细胞（hES细胞）是研究人类发育的宝贵工具，此外还在再生医学和药物研发中具有潜在应用价值。遗传背景和表观遗传影响在发育过程以及对药物和治疗等外部影响的反应中的作用已得到充分认识。印度次大陆存在巨大的种族多样性，这导致个体在药物反应和疾病易感性方面存在差异。基于这些原因，代表印度遗传多样性的新型hES细胞系对于组织分化、细胞功能研究以及药物反应特征的表征具有重要价值。我们从不适用于体外受精治疗的废弃III级人类胚胎的内细胞团（ICM）中获得了两个新的hES细胞系，BJNhem19和BJNhem20。人类白细胞抗原（HLA）同种型分析表明，它们在基因上与现有的hES细胞系不同。短串联重复序列（STR）分析表明，这两个细胞系源自同胞胚胎。这些细胞系在培养超过65代时呈现未分化表型，具有正常的核型，并表达多能性标志物，如TRA-1-60、TRA-1-81、阶段特异性胚胎抗原-4（SSEA-4）、碱性磷酸酶、DNMT3B、GABRB3、GDF3、OCT4、NANOG、SOX2、TERF1、TDGF、LEFTA、THY1和REX1。虽然这两个细胞系在体外均可分化为所有三个胚层的衍生物，但只有BJNhem20在移植到小鼠体内时能形成畸胎瘤。我们观察到，在无饲养层培养中，用二甲基亚砜（DMSO）诱导时，BJNhem20胚状体分化为心肌细胞的频率增加。从这种培养物中纯化的心肌细胞能够存活，并在培养中显示数周的节律性收缩。这些hES细胞系已被英国干细胞库接受保存，将成为国际干细胞界的有用资源。
