Secretion of oestrogen from murine-induced pluripotent stem cells co-cultured with ovarian granulosa cells in vitro
Secretion of oestrogen from murine-induced pluripotent stem cells co-cultured with ovarian granulosa cells in vitro
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DOI：
10.1042/cbi20100737
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发表时间：
2011-09-01
影响因子：
3.9
通讯作者：
Xu, Cong-Jian
中科院分区：
文献类型：
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作者：
Kang, Yu;Cheng, Ming-Jun;Xu, Cong-Jian
卵巢早衰(POF)是一种令人痛苦的状况，也是导致不孕的常见原因。目前还没有有效的治疗方法来克服生育能力的丧失。探索了一种从小鼠诱导的多能干细胞(MiPSCs)中提取雌激素的方法，作为治疗卵巢早衰的潜在方法。本研究将C57BL/6雌性小鼠注射孕马血清促性腺激素(PMSG)，获得卵巢颗粒细胞(GCs)，然后与miPSCs共培养。光镜下观察与GCs共培养的miPSCs的形态变化。免疫细胞化学和流式细胞仪检测卵泡刺激素受体(FSHR)表达。用放射免疫法测定培养上清液中雌二醇的含量。结果表明，表达FSHR的GCs占GCs的90%以上。GC组培养上清液中E-2浓度在第1天为62.4pg/ml，并呈时间依赖性下降。MPSCs-GC共培养组第1天E-2浓度为87.9pg/ml，第7天浓度为328.4 pg/ml，随着时间的延长，E-2浓度呈时间依赖性增加。我们的方法为研究miPSC的分化，特别是miPSC与GCs之间的相互作用提供了一种新的体外系统。这种方法的最终目标是在未来为POF提供一种治疗方法。
POF (premature ovarian failure) is a distressing condition that is a common cause of infertility. No effective treatment is available to overcome the loss of fertility. A method to derive oestrogen from miPSCs (mouse-induced pluripotent stem cells) was explored as a potential treatment for POF. In this study, C57BL/6 female mice were injected with PMSG (pregnant mare's serum gonadotropin) to obtain ovarian GCs (granulosa cells) and then co-cultured with miPSCs. The morphological changes in the miPSCs co-cultured with GCs were observed by light microscopy. The expression of FSHR (follicle-stimulating hormone receptor) was detected by immunocytochemistry and flow cytometry. Radioimmunoassay was used to analyse the level of E-2 (oestradiol) in culture supernatants. The results showed that the proportion of GCs expressing FSHR in GCs was over 90%. The E-2 concentration of the culture supernatant of the GC group was 62.4 pg/ml on day 1 and decreased in a time-dependent manner. The opposite situation was observed in the miPSCs-GC co-cultured group with an E-2 concentration of 87.9 pg/ml on day 1 that increased in a time-dependent manner to reach a concentration of 328.4 pg/ml on day 7. The data indicate that GC-like cells were effectively induced from miPSCs through indirect cell-to-cell contact. Our method provides a novel in vitro system to study miPSC differentiation, particularly the interactions between miPSCs and GCs. The ultimate goal of this approach would be to provide a treatment for POF in the future.