不同因子对花鲈胚胎干细胞增殖的影响

The effect of different factors on the proliferation of embryonic stem cells derived from Lateolabrax japonicus

胚胎干细胞(ES细胞)是从动物早期发育胚胎中分离出来的具有发育全能性的一种未分化细胞系。维持花鲈胚胎干细胞(LJES1)的体外生长、增殖及未分化状态需要在培养基中添加一些生长因子。实验通过配制省略1种或多种因子的培养基PESM0～10,计数LJES1细胞在各培养基中增殖的数量,确定各种生长因子对LJES1细胞的增殖作用。同时,对LIF因子和bFGF因子的作用进行了重点的研究,发现LIF因子对早期的LJES1细胞增殖几乎没有作用,其主要作用是维持LJES1细胞的未分化状态,但对晚期LJES1细胞的增殖有一定的作用;bFGF因子对LJES1细胞有强烈的刺激增殖的作用;鲈鱼胚胎抽提液(PEE)以及鲈鱼血清(FS)也促进了LJES1细胞的增殖,2-巯基乙醇(2-ME)具有还原血清中的含硫化合物、防止过氧化物对LJES1细胞的损害及促进贴壁的作用,因而也促进了LJES1细胞的增殖。

Embryonic stem cells(ES cells) are undifferentiated permanent cell line derived from inner cell mass cells and primordial germ cells of early developing embryos. Some growth factors were needed in culture medium to maintain LJES1 growth, proliferation and undifferentiation for a long time culture in vitro. Effects of various factors on the proliferation of LJES1 cells were examined by counting the number of multiplied cells in various mediums ( PESM0-10 ) omitting one or several factors. Furthermore, emphasis was focused on LIF and bFGF factor effects. LIF nearly had no effect on proliferation of early LJES1, just maintaining its undifferentiation, but it had a little effect on late LJES1;bFGF had a significant stimulative effect on the proliferation of LJES1.The number of LJES1 increased linearly at lower concentrations up to 2ng/ml bFGF, a nearly 2-3 fold increase was obtained,at higher than 2ng/ml concentration the proliferation-promoting effect was further enhanced but not significantly. Both sea perch embryo extract (PEE) and sea perch serum (FS) promoted the proliferation of LJES1,but their proliferating effect on LJES1 was not so significant as bFGF. The addition of 2-ME reduced the damage of peroxide on LJES1 cells, enhanced cell adhesion and promoted cell proliferation.