Apoptosis during mouse blastocyst formation: Evidence for a role for survival factors including transforming growth factor alpha
Apoptosis during mouse blastocyst formation: Evidence for a role for survival factors including transforming growth factor alpha
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DOI：
10.1095/biolreprod56.5.1088
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发表时间：
1997-05-01
影响因子：
3.6
通讯作者：
Schultz, RM
中科院分区：
文献类型：
--
作者：
Brison, DR;Schultz, RM
小鼠胚泡内细胞团（ICM）的细胞死亡是发育的正常特征，但很少有人知道这一事件是如何调节的，也不知道存活因子在植入前发育中可能发挥的作用。观察到生长因子，可以影响植入前发育，可以作为其他细胞类型的生存因子，使我们研究体外培养，培养过程中的胚胎密度，和转化生长因子α（TGF α）对胚泡细胞死亡的影响。在25 μ l KSOM +氨基酸培养基中从2-细胞阶段单独培养的小鼠囊胚显示出相对于体内形成的囊胚，细胞死亡的发生率（主要在ICM中）相似地增加至3倍。将胚胎培养密度增加到每25 μ l培养基中30个胚胎加速了发育，增加了最终囊胚细胞数，并部分（类似于50%）减少了体外培养诱导的细胞死亡增加。在单独培养的胚胎培养基中加入0.1 pM TGF α也部分（33%）降低了细胞死亡的增加，而没有加速发育或增加最终细胞数。在0.1 pM TGF α存在下培养分离的ICM 24 h也部分（33%）降低了细胞死亡的增加，全囊胚的末端脱氧核苷酸转移酶介导的dUTP缺口末端标记证实，如通过核酸内切酶激活所定义的，通过碎片化核检测到的细胞死亡是凋亡。这些实验的结果表明，内源性产生的生长因子可能作为细胞存活因子在植入前发育。
Mouse blastocysts undergo cell death in the inner cell mass (ICM) as a normal feature of development, but little is known as to how this event is regulated or as to the possible role of survival factors in preimplantation development. The observation that growth factors, which can influence preimplantation development, can act as survival factors in other cell types led us to investigate the effects of culture in vitro, embryo density during culture, and transforming growth factor alpha (TGF alpha) on cell death in the blastocyst. Mouse blastocysts cultured singly from the 2-cell stage in 25 mu I of medium KSOM + amino acids showed a similar to 3-fold increase in the incidence of cell death, predominantly in the ICM, relative to blastocysts formed in vivo. Increasing the density of embryo culture to 30 embryos per 25 mu I of culture medium accelerated development, increased final blastocyst cell number, and partially (similar to 50%) reduced the increase in cell death induced by culture in vitro. Addition of 0.1 pM TGF alpha to the medium of singly cultured embryos also partially (33%) reduced this increase in cell death without accelerating development or increasing final cell number. Culturing isolated ICMs for 24 h in the presence of 0.1 pM TGF alpha also partially (33%) reduced the increase in cell death, Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling of whole blastocysts confirmed that cell death as detected by fragmented nuclei was apoptotic, as defined by endonuclease activation. Results of these experiments suggest that endogenously produced growth factors may function as cell survival factors during preimplantation development.