Preparation of chicken myoblast cell line and regulation of gene expression related to the proliferation and differentiation of chicken myoblasts-Base line of control of meat production-

为了满足世界各地不断扩大的食品需求，可能需要改善肉类生产。本研究通过鸡成肌细胞系的建立和鸡成肌细胞增殖分化相关基因的表达调控，为建立高效的肉品生产体系奠定了基础。鸡成肌细胞系的制备。鸡成肌细胞可以通过克隆形成程序克隆，但几代都不能保持增殖。B.将SV40 Large T基因导入鸡原代成肌细胞，增强其增殖活性。细胞克隆中瞬时出现SV40大T的表达。克隆鸡肌肉细胞特异性蛋白启动子序列，将其连接到绿色荧光蛋白表达载体pd2EGFP-1上，并将其导入鸡成肌细胞。分析目前仍在继续，准备工作在…结论：1.利用该方法可获得更多的鸡肌肉细胞株。鸡成肌细胞增殖分化调控因子的筛选。在鸡成肌细胞培养中，MyoD和Mygenin的mRNA表达分别在12小时和2小时达到高峰，之后逐渐下降。加入前列腺素E_2(PGE_2)后，肌球蛋白、CPK活性和myd基因表达增加，提示PGE_2是一种刺激肌细胞分化的因子。在心肌细胞培养中加入酮体--β-羟基丁酸酯(BTB)可促进肌细胞生长期的增殖和终末分化时成肌细胞的融合，这些作用可被培养液中的能量来源所改变。孵化后的雏鸡腹部残留的蛋黄似乎含有丰富的代谢功能的蛋白质。低浓度(1~100 ug)对肌细胞分化无影响，高浓度(1~3 mg)则诱导细胞死亡。较少

The improvement of meat production may be requested to meet the expanding demands for foods all over the world. In the present study, preparation of chicken myoblast cell line and regulation of gene expression related to the proliferation and differentiation of chicken myoblasts was conducted to construct the base line for promoting the efficient meat production system.1. Preparation of chicken myoblast cell linea. Chicken myoblasts could be cloned by the colony formation procedure, while the proliferation could not be maintained for several generations.b. b. SV40 Large T was transfected to chicken primary myoblasts for potentiating the proliferation activity. The expression of SV40 Large T was transiently appeared in the cell clone.c. Promoter sequences of specific proteins in chicken muscle cells were subjected for cloning and ligated to plasmid (pd2EGFP-1) which expressed GFP, and then transfected to the chicken primary myoblasts. The analysis is currently continued and the preparat … More ion of chicken muscle cell line is scheduled with aid of this procedure.2. Screening of factors regulating proliferation and differentiation in chicken myoblastsa. In the chicken myoblast culture, MyoD and Myogenin mRNA expression were peaked at 12 and 2h hr, respectively and they were followed by gradual decrease.b. Myosin, CPK activity and MyD mRNA expression increased by addition of prostaglandin E2 (PGE2), suggesting that PGE2 is a factor stimulating the differentiation of muscle cells.c. Proliferation in the myocyte growth stage and fusion of myoblasts in the terminal differentiation was stimulated by addition of beta-hydroxybutyrate (BTB), a ketone body, in the myocyte culture and these effects were modified by the amount of energy source present in the medium.d. Residual yolk present in the abdomen of post-hatched chicks appears to contain a plenty of proteins rich in metabolic function. Addition of the proteins extracted from the residual yolk was no influence on the muscle cell differentiation at the low concentration (1-100 ug) but induced the cell death at the high concentration (1 or 3 mg). Less