Establishment of SSEA-1-and Oct-4-expressing rat embryonic stem-like cell lines and effects of cytokines of the IL-6 family on clonal growth
Establishment of SSEA-1-and Oct-4-expressing rat embryonic stem-like cell lines and effects of cytokines of the IL-6 family on clonal growth
复制标题
DOI：
10.1006/excr.2000.4940
复制
发表时间：
2000-08-01
影响因子：
3.7
通讯作者：
Wobus, AM
中科院分区：
文献类型：
--
作者：
Vassilieva, S;Guan, KM;Wobus, AM
在这里，我们证明了长期培养碱性磷酸酶阳性大鼠胚胎干细胞样（RES）细胞系。RES细胞的特征在于其典型的高度紧凑的细胞簇中的生长，这被发现是对酶促解离敏感。RES细胞表达阶段特异性胚胎抗原-1（SSEA-1）和转录因子Oct-4，但Oct-4 mRNA的检测水平低于小鼠ES细胞。一旦建立组织培养，RES细胞能够在克隆条件下在不存在饲养细胞的情况下生长。细胞因子：在克隆测定中，比较分析了已知维持小鼠ES细胞未分化状态的白细胞介素-g家族中的两种细胞系RES-1和RES-15维持未分化生长的能力。大鼠睫状神经营养因子（rCNTF）、人抑瘤素M（hOSM）、白细胞介素6（IL-6）和可溶性白细胞介素6受体（IL-6/sIL-6 R）均能支持RES细胞的克隆性生长，但其作用不及白血病抑制因子（LIF）。当RES-1和RES-15细胞在没有饲养细胞的情况下培养时，SSEA-1表达在LIF和LIF + rCNTF存在下分别在克隆生长后维持。Oct-4 mRNA在RES-15细胞中显著检测到，当在不存在饲养细胞的情况下在用LIF和/或IL-6/sIL-6 R替代的培养基中培养时，以及在没有细胞因子的情况下。总之，大鼠胚胎干细胞样细胞系可以建立从大鼠囊胚，并能够增殖为未分化的碱性磷酸酶阳性细胞。胚胎干细胞的特性，如SSEA-1和Oct-4的表达，由IL-6家族的细胞因子维持，但最重要的是由LIF维持。(C)北京大学出版社.
Here, we demonstrate long-term cultivation of alkaline phosphatase-positive rat embryonic stem-like (RES) cell lines. RES cells were characterized by their typical growth in highly compacted cell clusters, which were found to be sensitive against enzymatic dissociation. RES cells expressed stage-specific embryonic antigen-1 (SSEA-1) and transcription factor Oct-4, but Oct-4 mRNA was detected at lower levels compared to mouse ES cells. Once established to tissue culture, RES cells were able to grow in the absence of feeder cells under clonal conditions. Cytokines: of the interleukin-g family known to maintain the undifferentiated state of mouse ES cells were comparatively analyzed for their capacity to maintain the undifferentiated growth of two cell lines, RES-1 and RES-15, in a clonal assay. Rat ciliary neurotrophic factor (rCNTF), human oncostatin M (hOSM), and interleukin-6 and soluble interleukin-6 receptor (IL-6/sIL-6R) were found to support clonal growth of RES cells, but the cytokines did not reach the efficiency of the colony forming ability of leukemia inhibitory factor (LIF). When RES-1 and RES-15 cells were cultivated without feeder cells, SSEA-1 expression was maintained after clonal growth in the presence of LIF and LIF + rCNTF, respectively. Oct-4 mRNA was significantly detected in RES-15 cells when cultivated in the absence of feeder cells in media substituted by LIF and/or IL-6/sIL-6R, as well as without cytokines. In summary, rat embryonic stem-like cell lines could be established from rat blastocysts and were able to proliferate as undifferentiated alkaline phosphatase-positive cells. Embryonal stem cell properties, such as SSEA-1 and Oct-4 expression, were maintained by members of the IL-6 family of cytokines, but most significantly by LIF. (C) 2000 Academic Press.