豬心臟纖維母細胞之分離、鑑別與膠原蛋白基因之轉錄

Isolation, Characterization and Transcription of Collagen Genes of Porcine Cardiac Fibroblasts

黃琇琴(Hsiu-Chin Huang) ； 李文權(Wen-Chuan Lee) ； 鄭益謙(Ivan-Chen Cheng) ； 楊平政(Ping-Cheng Yang)

《中華民國獸醫學會雜誌》 22卷4期 (1996/08) Pp. 281-292

https://doi.org/10.7009/JTCSVS.199608.0281

摘要

Fibroblasts were isolated by collagenase digestion techniques from 80 days embryonic porcine heart. The cells were subcultured every 5 to 7 days. The doubling time of the cells with eight and twelve passages approximated five and four days, respectively. The result from immunofluorescence staining using vimentin, α-smooth muscle actin and desmin monoclonal antibodies indicated porcine embryonic fibroblasts presenting typical morphology of vimentin-containing intermediate filaments. We further used this cell line to study the effect of growth factors, including aFGF, bFGF and TGF-β, on the expression of type I and type III collagen genes by RT-PCR method. The data showed that the expression of type I collagen gene was increased by 1.2-fold in cells treated with10 ng/mL TGF-βwhen compared to the control cells(P＜0.05). In contrast, there was no significant difference in the mRNA levels among the cells treated with aFGF and bFGF at any dosage. The above observations indicated that we have successfully maintained the cardiac fibroblasts from porcine embryos and demonstrated that the type I collagen gene expression in these cells is stimulated by TGF-β. We suggest that the cell line can be extensively used as a model to study the pathogenesis of cardiovascular diseases such as hypertrophic cardiomyopathy.

關鍵字

豬 ； 心臟纖維母細胞 ； 生長因子 ； 膠原蛋白

並列摘要

Fibroblasts were isolated by collagenase digestion techniques from 80 days embryonic porcine heart. The cells were subcultured every 5 to 7 days. The doubling time of the cells with eight and twelve passages approximated five and four days, respectively. The result from immunofluorescence staining using vimentin, α-smooth muscle actin and desmin monoclonal antibodies indicated porcine embryonic fibroblasts presenting typical morphology of vimentin-containing intermediate filaments. We further used this cell line to study the effect of growth factors, including aFGF, bFGF and TGF-β, on the expression of type I and type III collagen genes by RT-PCR method. The data showed that the expression of type I collagen gene was increased by 1.2-fold in cells treated with10 ng/mL TGF-βwhen compared to the control cells(P＜0.05). In contrast, there was no significant difference in the mRNA levels among the cells treated with aFGF and bFGF at any dosage. The above observations indicated that we have successfully maintained the cardiac fibroblasts from porcine embryos and demonstrated that the type I collagen gene expression in these cells is stimulated by TGF-β. We suggest that the cell line can be extensively used as a model to study the pathogenesis of cardiovascular diseases such as hypertrophic cardiomyopathy.

並列關鍵字

Porcine ； Cardiac fibroblasts ； Growth actors ； Collagen