Growth factors sustain primordial germ cell survival, proliferation and entering into meiosis in the absence of somatic cells
Growth factors sustain primordial germ cell survival, proliferation and entering into meiosis in the absence of somatic cells
复制标题
DOI：
10.1016/j.ydbio.2005.06.036
复制
发表时间：
2005-09-01
影响因子：
2.7
通讯作者：
De Felici, M
中科院分区：
文献类型：
--
作者：
Farini, D;Scaldaferri, ML;De Felici, M
已知哺乳动物的原始生殖细胞（PGCs），即卵母细胞和精原细胞的前体，其生存和增殖依赖于特定的生长因子以及其他尚未确定的化合物。据信，与邻近体细胞的黏附对于防止PGCs在失去适当的细胞间接触时发生凋亡也至关重要。这就解释了为什么在没有合适的能产生可溶性因子并表达防止PGCs凋亡和刺激其增殖所必需的表面分子的细胞饲养层的情况下，目前无法将分离的小鼠PGCs在培养中维持超过几个小时。在本文中，我们确定了一组可溶性生长因子的组合，即KL、LIF、BMP - 4、SDF - 1、bFGF以及一些化合物（N - 乙酰 - L - 半胱氨酸、毛喉素、视黄酸），它们能够在没有体细胞支持的情况下维持小鼠PGCs的生存和自我更新。我们表明，在允许PGCs黏附于无细胞基质的培养条件下，这些生长因子和化合物能够在2天内防止PGCs发生显著水平的凋亡，刺激它们的增殖，并且当组合中省略LIF时，能使大多数PGCs进入减数分裂前期I并进展。这些结果首次使得在没有体细胞支持的情况下为纯化的哺乳动物PGCs建立培养条件成为可能，并且应该会使控制这类对早期配子发生至关重要的细胞发育过程的分子解析变得更容易。（c）2005年爱思唯尔公司。保留所有权利。
It is known that mammalian primordial germ cells (PGCs), the precursors of oocytes and prospermatogonia, depend for survival and proliferation on specific growth factors and other undetermined compounds. Adhesion to neighboring somatic cells is also believed to be crucial for preventing PGC apoptosis occurring when they loss appropriate cell to cell contacts. This explains the current impossibility to maintain isolated mouse PGCs in culture for periods longer than a few hours in the absence of suitable cell feeder layers producing soluble factors and expressing surface molecules necessary for preventing PGC apoptosis and stimulating their proliferation.In the present paper, we identified a cocktail of soluble growth factors, namely KL, LIF, BMP-4, SDF-1, bFGF and compounds (N-acetyl-L-cysteine, forskolin, retinoic acid) able to sustain the survival and self-renewal of mouse PGCs in the absence of somatic cell support. We show that under culture conditions allowing PGC adhesion to an acellular substrate, such growth factors and compounds were able to prevent the occurrence of significant levels of apoptosis in PGCs for 2 days, stimulate their proliferation and, when LIF was omitted from the cocktail, allow most of them to enter into and progress through meiotic prophase I. These results consent for the first time to establish culture conditions for purified mammalian PGCs in the absence of somatic cell support and should make easier the molecular dissection of the processes governing the development of such cells crucial for early gametogenesis. (c) 2005 Elsevier Inc. All rights reserved.