Department of Stem Cell and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Arch Iran Med. 2011 Mar;14(2):96-103.
Despite the suitability of a mouse model for preclinical investigations; little is known regarding mesenchymal stem cells derived from murine amniotic fluid. This is the subject of the present study.
Amniotic fluid was collected from NMRI mice during the second weeks of pregnancy and plated. The cells that adhered to the culture surfaces were propagated with three successive subcultures and then characterized. To determine the differentiation potential, the cells were cultivated under osteogenic, adipogenic, and chondrogenic conditions, and followed by specific staining and RT-PCR analysis for differentiation. The proliferative potential of the cells were measured with clonogenic assays, population doubling time and number and by growth curve plotting. Cellular aging was investigated with the senescence-associated ß-galactosidase staining method.
The amniotic fluid primary cell culture was composed of round flattened and fibroblastic cells. The latter dominated the culture after several passages. Successful tripotent differentiation of the isolated cells into bone, cartilage and adipose cells were indicative of their mesenchymal stem cells nature. The isolated cells appeared to be relatively proliferative cells as confirmed by the population doubling time value which was equal to about 69 hours. Furthermore, the cells were relatively clonogenic and they tended to initiate proliferation immediately after plating (there was no lag phase in their growth curve). ß-galactosidase positive cells were first observed at passage 3 and increased in number with subsequent passages.
Collectively it was concluded that murine amniotic fluid contained mesenchymal stem cells with relatively high proliferation property and typical tripotent differentiation potential.
尽管小鼠模型适合进行临床前研究，但对于来源于鼠类羊水的间充质干细胞，人们知之甚少。本研究即针对这一主题展开。
在妊娠第 2 周时从 NMRI 小鼠中采集羊水并进行铺板。贴壁的细胞经 3 次连续传代进行增殖，并对其进行特性鉴定。为了确定分化潜能，将细胞在成骨、成脂和成软骨条件下培养，随后进行特定染色和 RT-PCR 分析以进行分化鉴定。通过集落形成实验、群体倍增时间、细胞数量和生长曲线绘制来测量细胞的增殖潜能。通过衰老相关的β-半乳糖苷酶染色方法研究细胞衰老。
羊水原代细胞培养物由圆形扁平细胞和纤维母细胞组成。后者在传代几次后成为培养物中的主要细胞。分离细胞成功地三向分化为骨、软骨和脂肪细胞，这表明它们具有间充质干细胞的特性。分离细胞似乎是相对增殖的细胞，因为其群体倍增时间值约为 69 小时。此外，细胞具有相对的集落形成能力，它们在接种后立即开始增殖（生长曲线中没有停滞期）。β-半乳糖苷酶阳性细胞在第 3 代时首次观察到，并随着传代次数的增加而增加。
综上所述，鼠类羊水含有具有相对较高增殖特性和典型三向分化潜能的间充质干细胞。
