Liu Te, Huang Yongyi, Bu Yanzhen, Zhao Yanhui, Zou Gang, Liu Zhixue
Shanghai Geriatric Institute of Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200031, P.R. China.
School of Life Science and Technology, Tongji University, Shanghai 200092, P.R. China.
Mol Med Rep. 2014 Jul;10(1):21-8. doi: 10.3892/mmr.2014.2199. Epub 2014 Apr 29.
Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin- HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and β-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA‑E‑cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and β-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells.
多能性人羊水细胞（HuAFCs）在体外可分化为多种类型的体细胞。然而，据我们所知，它们从未被描述过可分化为卵母细胞样细胞。在本研究中，通过在含有牛卵泡液和β-巯基乙醇的培养基中培养，诱导E-钙黏蛋白阳性和E-钙黏蛋白阴性HuAFC亚群分化为卵母细胞样细胞。E-钙黏蛋白阳性HuAFCs高表达DAZL。诱导后，在E-钙黏蛋白阳性HuAFCs中发现了具有卵母细胞样表型的细胞，这些细胞表达生殖细胞和卵母细胞特异性标志物（VASA、ZP3和GDF9）以及减数分裂标志物（DMC1和SCP3）。当使用特异性小干扰RNA（siRNA）抑制E-钙黏蛋白阳性HuAFCs中的E-钙黏蛋白时，DAZL表达水平降低。诱导后，与siRNA对照HuAFCs相比，siRNA-E-钙黏蛋白HuAFCs的形态较差，生殖细胞特异性标志物的表达水平较低。因此，E-钙黏蛋白阳性HuAFCs更容易被牛卵泡液和β-巯基乙醇诱导分化为卵母细胞样细胞。此外，E-钙黏蛋白阳性HuAFCs表现出DAZL蛋白表达的潜在特征，因此推测牛卵泡液作用于DAZL蛋白并促进E-钙黏蛋白阳性HuAFCs分化为卵母细胞样细胞。
