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鸡胚脊髓细胞在无血清培养中的存活及神经突生长

Survival and neurite growth of chick embryo spinal cord cells in serum-free culture.

作者信息

Tanaka H, Obata K

出版信息

Brain Res. 1982 Jul;256(3):313-21. doi: 10.1016/0165-3806(82)90143-2.

Abstract

Cell survival and neurite growth were investigated in serum-free spinal cord cell cultures on polyornithine coating (PORN). Cells were obtained from 6- or 7-day-old chick embryos. Isolated spinal cord cells required promoting factors for their survival and neurite growth. The survival-promoting factors were initially present in spinal cord cells. High density cultures, co-cultures with spinal cord explants, and spinal cord extract promoted survival of isolated spinal cord cells in MEM with no additives. Other tissue extracts (brain, liver, heart and skeletal muscle), serum, and serum-free conditioned medium (SF-CM) of muscle or glioma C6 cells also promoted survival. The active substances in the brain extract and SF-CM were shown to be protein and were separated into 3 fractions (approximately molecular weight 150,000, 70,000, 40,000) by gel filtration chromatography. Survival and neurite growth were suggested to be promoted by different factors because: (1) survival was promoted by both tissue extract and SF-CM, but neurite growth was promoted only by SF-CM; (2) the neurite growth-stimulating activity of SF-CM was lost following dialysis and heat (100 degrees C, 2 min) treatment; however, the survival-promoting activity was not. It was also suggested that spinal cord cells produce neurite growth promoting factors, but did not initially contain these factors.

摘要

在聚鸟氨酸包被(PORN)的无血清脊髓细胞培养物中研究细胞存活和神经突生长。细胞取自6或7日龄鸡胚。分离的脊髓细胞的存活和神经突生长需要促生长因子。促存活因子最初存在于脊髓细胞中。高密度培养、与脊髓外植体共培养以及脊髓提取物可促进MEM中无添加剂的分离脊髓细胞的存活。其他组织提取物(脑、肝、心脏和骨骼肌)、血清以及肌肉或胶质瘤C6细胞的无血清条件培养基(SF-CM)也可促进存活。脑提取物和SF-CM中的活性物质被证明是蛋白质,并通过凝胶过滤色谱法分离成3个组分(分子量约为150,000、70,000、40,000)。细胞存活和神经突生长被认为是由不同因子促进的,原因如下:(1)组织提取物和SF-CM均可促进存活,但只有SF-CM可促进神经突生长;(2)SF-CM的神经突生长刺激活性在透析和加热(100℃,2分钟)处理后丧失;然而,促存活活性并未丧失。还表明脊髓细胞产生神经突生长促进因子,但最初并不含有这些因子。

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