Alikani Mina, Munné Santiago
Tyho-Galileo Research Laboratories, West Orange, NJ 07052, USA.
Stem Cell Rev. 2005 Dec;1(4):337-43. doi: 10.1385/SCR:1:4:337.
Human embryonic stem cells are derived from the inner cell mass of the human blastocyst. Presumably normal (frozen/thawed) human preimplantation embryos that remain unused following assisted reproduction procedures have provided the main source of blastocysts for stem cell derivation. Alternatively, embryos have been generated from gametes donated for the unique purpose of in vitro fertilization, blastocyst culture, and stem cell isolation. This article describes two previously published methods--and the background to those methods--that allow the use of nonviable embryos excluded from transfer and cryopreservation as a source of stem cells. The first method is based on the observation that some blastomeres from embryos with abnormal division during the first 3-5 d in culture can continue very limited development in isolation. When aggregated in a chimaeric form, some of these blastomeres can contribute to the formation of normally organized blastocysts. Blastocysts so obtained provide a route to embryonic stem cells from otherwise nonviable embryos. Thus the inner cell masses of blastocysts obtained from trisomic embryos were placed on feeder cells and cultured for seven additional days, following which the resulting cell colonies were examined for chromosome content. The second method concerns embryos diagnosed with specific chromosome abnormalities many of which are incompatible with life. Some of these aneuploidies do not preclude development to the blastocyst stage in culture. A proportion of these cells were found to be disomic and the cultures were shown to express OCT-4, a molecular marker for pluripotent cells. This apparent correction of the trisomic state in some cells within the colonies suggests that embryos with cromosomal abnormalities incompatible with life may be another source of human embryonic stem cells.
人类胚胎干细胞源自人类囊胚的内细胞团。辅助生殖程序后剩余未使用的推测正常(冷冻/解冻)的人类植入前胚胎是干细胞衍生所用囊胚的主要来源。另外,胚胎由为体外受精、囊胚培养和干细胞分离这一独特目的而捐赠的配子产生。本文描述了两种先前发表的方法——以及这些方法的背景——这些方法允许将被排除在移植和冷冻保存之外的无活力胚胎用作干细胞来源。第一种方法基于这样的观察结果:在培养的前3 - 5天内分裂异常的胚胎中的一些卵裂球在分离后可以继续非常有限地发育。当以嵌合形式聚集时,其中一些卵裂球可以促成正常组织的囊胚的形成。如此获得的囊胚为从其他方面无活力的胚胎获得胚胎干细胞提供了一条途径。因此,将从三体胚胎获得的囊胚的内细胞团置于饲养细胞上并再培养7天,之后检查所得细胞集落的染色体含量。第二种方法涉及被诊断患有特定染色体异常的胚胎,其中许多与生命不相容。其中一些非整倍体并不排除在培养中发育到囊胚阶段。发现这些细胞中有一部分是二体的,并且培养物显示表达OCT - 4,这是一种多能细胞的分子标记。集落内一些细胞中三体状态的这种明显纠正表明,患有与生命不相容的染色体异常的胚胎可能是人类胚胎干细胞的另一个来源。