DIFFERENTIATION IN-VITRO OF MOUSE EMBRYOS TO STAGE OF EARLY SOMITE
DIFFERENTIATION IN-VITRO OF MOUSE EMBRYOS TO STAGE OF EARLY SOMITE
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DOI：
10.1016/0012-1606(73)90145-0
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发表时间：
1973-01-01
影响因子：
2.7
通讯作者：
HSU, YC
中科院分区：
文献类型：
--
作者：
HSU, YC
以重组鼠尾胶原为贴壁基质，小鼠囊胚体外连续分化为体节早期。要做到这一点，似乎必须克服两个差异化障碍。首先，从内细胞团形成卵柱，可以通过在加热灭活的小牛血清中孵化胚胎来克服。第二种是从前卵子阶段形成的早期体节，可以用人脐带血清代替胎牛血清来克服。小鼠囊胚最初在Eagle‘s最低基本培养液中与小牛血清孵育。剥离透明带后，裸露的胚泡平铺在胶原表面。此后不久，滋养层细胞侵入下面的胶原蛋白，留下圆形的内细胞团从胶原蛋白表面突出。用胎牛血清代替培养液中的小牛血清，内细胞团分化为内胚层和外胚层，形成卵柱，卵柱迅速伸长，形成胚外和胚胎区。然而，从这一点开始，胚胎区域在胎牛血清中收缩，由此形成的卵黄囊不包含胚胎本身。当在卵筒阶段(相当于妊娠7.5天的胚胎)末期用人脐带血清代替胎牛血清时，神经组织、心腔和体节形成。
Mouse blastocysts continuously differentiatein vitroto the early somite stage with reconstituted rat tail collagen as the substrate for the attachment. In order for this to occur, it appears that two differentiation barriers must be overcome. The first, the formation of egg cylinders from the inner cell mass, can be overcome by incubating embryos in heat-inactivated fetal calf serum. The second, the formation of the early somite from the presomite stage, can be overcome by replacing fetal calf serum with human cord serum.Mouse blastocysts were initially incubated with calf serum in Eagle's minimum essential medium. After shedding the zona pellucida, the denuded blastocysts lay flat on the surface of the collagen. Soon thereafter, trophoblastic cells invaded the underlying collagen leaving the rounded inner cell mass protruding from the surface of the collagen. By replacing calf serum in the medium with fetal calf serum the inner cell mass differentiated into endoderm and ectoderm to form an egg cylinder.The egg cylinder rapidly became elongated and formed extraembryonic and embryonic regions. However, the embryonic region shrank from this point on in the fetal calf serum, and the resulting yolk sac formation did not contain the embryo proper. When fetal calf serum was replaced with human cord serum at the end of the egg cylinder stage (equivalent to embryos of about 7.5 days gestation) neural tissue, cardiac chambers, and somites were formed.