The mitotic manipulation of cytotrophoblast differentiation in vitro
The mitotic manipulation of cytotrophoblast differentiation in vitro
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DOI：
10.1016/j.placenta.2006.05.009
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发表时间：
2007-05-01
期刊：
影响因子：
3.8
通讯作者：
Baker, P. N.
中科院分区：
文献类型：
--
作者：
Crocker, I. P.;Arthur, P.;Baker, P. N.
胎盘合体滋养层细胞在优化胎儿与母体的相互作用中起着至关重要的作用。合体滋养层细胞是由底层细胞滋养层细胞分化融合而成。这一过程在复杂的妊娠中是异常的。我们假设细胞周期退出决定了细胞滋养层细胞的表型决策。因此，我们研究了广谱有丝分裂抑制剂对细胞滋养层细胞分化的影响。从正常妊娠足月胎盘中分离绒毛组织，并在Netwell支架上培养。48h后，原合体滋养层细胞脱落，下层细胞滋养层暴露。用有丝分裂阻滞剂(阿糖胞苷、秋水仙胺、环己胺、盐酸阿霉素、羟基脲、L-米莫辛、紫藜芦醇A)处理绒毛。回收培养基作乳酸脱氢酶(LDH)和人绒毛膜促性腺激素(HCG)检测，分别检测组织活性和细胞滋养细胞分化。通过Ki-67免疫识别对所得到的组织进行增殖活性处理。Colcemid、环己胺、羟基脲和紫杉醇A在48it孵育后显示出显著的细胞毒性。0.01 mM和0.1 mM阿糖胞苷、阿霉素和L-米莫辛对绒毛组织释放的乳酸脱氢酶无明显影响。随着细胞滋养层细胞的分化，人绒毛膜促性腺激素的产生呈指数增加。较高浓度的Ara-C和L-米莫辛显著促进hCG的产生。此外，阿糖胞苷和L含羞草抑制细胞和细胞滋养层细胞的增殖，抑制细胞的DNA合成和复制，抑制绒毛成分的活跃增殖，夸大细胞滋养层细胞的生化分化。因此，细胞周期中断是细胞滋养层细胞分化的基本触发因素。这种方法提供了一种鼓励合体滋养层细胞形成的机制，并可能对合体滋养层覆盖减弱的情况有好处。(C)2006爱思唯尔有限公司。保留所有权利。
The placental syncytiotrophoblast is of paramount importance in optimising feto-maternal interactions. Syncytiotrophoblast is generated by the differentiation and fusion of underlying cytotrophoblasts. This process is aberrant in complicated pregnancies. We hypothesized that cell cycle withdrawal determines the phenotypic decision-making of cytotrophoblasts. We therefore investigated the effects of broad-spectrum mitotic inhibitors on cytotrophoblast differentiation.Villous tissue was dissected from term placentae of normal pregnancies and cultured on Netwell supports. Over 48 h, the original syncytiotrophoblast was detached and underlying cytotrophoblasts exposed. The resulting villi were treated with mitotic blockers (Ara-C, colcemid, cyclohexamide, doxorubicin hydrochloride, hydroxyurea, L-Mimosine, purvalanol A). The media was recovered and analysed for lactacte dehydrogenase (LDH) and human chorionic gondadotrophin (hCG), markers of tissue viability and cytotrophoblast differentiation, respectively. The resulting tissue was processed for proliferative activity thorough Ki-67 immunorecognition.Colcemid, cyclohexamide, hydroxyurea, and purvalanol A showed significant cytotoxicity over 48 It incubation. Villous tissue exposed to 0.01 mM and 0.1 mM Ara-C, doxorubicin hydrochloride and L-Mimosine showed no increase in liberated LDH. hCG production increased exponentially with cytotrophoblast differentiation. Higher concentrations of Ara-C and L-Mimosine significantly encouraged hCG production. In addition, total cell and cytotrophoblast proliferation were reduced with Ara-C and L-Mimosine treatment.The inhibition of DNA synthesis and replication with Ara-C and L-Mimosine suppressed active proliferation of villus components and exaggerated the biochemical differentiation of cytotrophoblasts. Cell cycle disruption is therefore a basic trigger for cytotrophoblast differentiation. This approach provides a mechanism for encouraging syncytiotrophoblast formation and may hold benefits for conditions where syncytiotrophoblast cover is attenuated. (c) 2006 Elsevier Ltd. All rights reserved.