血清体积分数对培养黑山羊成纤维细胞的影响

Effect of Different Serum Volume Fraction on Cultured Fibroblasts from Black Goat

为了建立并完善吕梁黑山羊耳缘成纤维细胞培养体系,为进一步的克隆试验提供最佳生长状态的供体细胞,在常规培养液的基础上添加不同浓度的新生牛血清(0,5%,10%,20%,40%),每天在倒置显微镜下观察培养细胞的生长状态,并用台盼蓝染色法统计细胞数目,连续观察7 d,绘制生长曲线,比较不同血清体积分数的培养液对吕梁黑山羊耳缘成纤维细胞体外培养的影响。结果表明,吕梁黑山羊耳缘成纤维细胞虽然在不同血清体积分数的5组中均有生长增殖,但生长状态不同,差距较大;其中,10%的血清培养液试验组培养的细胞生长状况最好,细胞形态完整清晰,增殖快,数量多,更适合此细胞的生长。不同体积分数血清的培养液对成纤维细胞的生长具有一定影响,10%血清体积分数的培养液最适合吕梁黑山羊耳缘成纤维细胞的培养。结果可为进一步研究克隆、基因转染、构建基因文库等提供生物学材料。

To establish and perfect the fibroblast culture system, to provide optimal growth state of donor cells for further cloning experiments, in this paper, we added different volume fraction of new bovine serum（0, 5%, 10%, 20%, 40%）on the basis of conventional cell culture medium, the growth state and the number of cultured cells was counted by trypan blue staining and observed under inverted microscope daily. We continued observation for seven days, then drew the growth curve, compared the influence of different serum volume fraction in nutrient solution to in vitro culture fibroblasts from Lüliang native goat ear marginal cell. The results showed that although the growth and proliferation of ear marginal fibroblasts from Lüliang black goat were found in the 5 groups, the growth condition had significant disparity with different newborn serum volume fraction. Among them, the growth of the cells with the 10% serum volume fraction was the best. In this case, the cell morphology was complete and clear, the proliferation was fast, and the number was large, which was more suitable for the growth of this cell. The results indicate that different volume fraction of serum culture has certain influence on the growth of fibroblasts. The culture solution of 10% serum volume fraction is the most suitable for primary culture of the fibroblasts from Lüliang native goat ear margin cells. This result can provide biological materials for further study on gene cloning, gene transfection, build a gene library, etc..