STIMULATION OF EARLY EMBRYONIC-DEVELOPMENT IN THE SHEEP BY COCULTURE WITH OVIDUCT EPITHELIAL-CELLS
STIMULATION OF EARLY EMBRYONIC-DEVELOPMENT IN THE SHEEP BY COCULTURE WITH OVIDUCT EPITHELIAL-CELLS
复制标题
DOI：
10.1530/jrf.0.0810023
复制
发表时间：
1987-09-01
期刊：
影响因子：
--
通讯作者：
MOOR, RM
中科院分区：
文献类型：
--
作者：
GANDOLFI, F;MOOR, RM
为探讨体细胞支持对绵羊受精卵卵裂率和存活率的影响，将434枚原核卵与输卵管细胞或成纤维细胞共培养3~6天，并将77枚原核卵单独在培养液中培养。在培养的前3天，95%的单细胞卵在任一饲养层上规则地分裂为未压实的桑椹，但只有13%的单细胞卵在单独的培养液中经历了类似的规则卵裂。尽管共培养组的卵裂率相同，但从移植给受体动物后正常发育的能力来看，只有33%的胚胎生长在成纤维细胞上，而80%的胚胎生长在输卵管细胞上。输卵管组的胚胎存活率与供体绵羊桑椹胚直接移植给受体绵羊后的胚胎存活率相当。与输卵管细胞共培养6d后，42%的胚胎发育为扩张囊胚，而与成纤维细胞共培养的胚胎仅有4.5%发育为扩张囊胚。在两个共培养组中，几乎所有剩余的胚胎都在第四次卵裂时被阻断。在输卵管细胞上移植原核期的囊胚，30%的囊胚存活。我们得出的结论是：(1)在受精后的前3天，卵裂在不同的饲养层上将以正常的速度进行，但输卵管细胞似乎是获得完全胚胎活力所必需的；(2)在我们的系统中，输卵管细胞能够支持胚胎通过关键的第4细胞周期，而成纤维细胞几乎完全不能支持这一关键时期；以及(3)很明显，囊胚的形态形成所需的因素可能不足或不同于赋予其后续发育能力的因素。
To examine the effects of somatic cell support on the cleavage and viability of fertilized sheep eggs, 434 pronucleate eggs were co-cultured for 3 or 6 days on oviduct cells or fibroblasts and 77 eggs were cultured in medium alone. During the first 3 days in culture 95% of the single-celled eggs cleaved regularly to non-compacted morulae on either of the feeder-layers but only 13% underwent similar regular cleavage in medium alone. Despite the identical cleavage rates in the co-culture groups, only 33% of embryos grown on fibroblasts as compared with 80% of embryos grown on oviduct cells were fully viable as judged by their ability to develop normally after transfer to recipient animals. The viability of embryos in the oviduct group was equal to that obtained after the direct transfer of morulae from donor to recipient sheep. After 6 days in culture 42% of embryos co-cultured with oviduct cells developed into expanded blastocysts as compared with only 4.5% cultured on fibroblasts. In both co-culture groups virtually all the remaining embryos blocked during the 4th cleavage. When transferred, 30% of blastocysts grown from the pronucleate stage on oviduct cells were viable. We conclude that: (1) during the first 3 days after fertilization cleavage will progress at a normal rate on different feeder-layers but oviduct cells appear to be required for the acquisition of full embryonic viability; (2) in our system oviduct cells are able to support passage of embryos through the critical 4th cell cycle, whilst fibroblasts are almost entirely unable to support this critical phase; and (3) it is apparent that the factors necessary for the morphological formation of the blastocyst may be insufficient or different from those which endow it with subsequent developmental ability.