奶牛乳腺上皮细胞的二维和三维培养及培养后酪蛋白基因的表达

Two-and Three-Dimensional Cultivation of Bovine Mammary Epithelial Cells and Gene Expression of Casein after Cultured

本试验研究了二维和三维培养对奶牛乳腺上皮细胞(BMEC)形态与β-酪蛋白、κ-酪蛋白、αs1-酪蛋白基因表达的影响。试验采用内蒙古地区3头5岁泌乳期健康荷斯坦奶牛的乳腺组织,将BMEC经3代培养纯化后分别进行二维和三维培养,观察不同培养模式下细胞的生长形态,测定β-酪蛋白、κ-酪蛋白、αs1-酪蛋白基因的表达量。结果表明:2种培养模式下的BMEC呈现不同的细胞形态,二维培养的BMEC呈典型铺路石样细胞形态,三维培养的BM EC则形成了类腺泡的聚集物和管腔结构。利用三维培养的BM EC的αs1-酪蛋白基因的表达量显著地高于二维培养(P<0.05),三维培养的BMEC的β-酪蛋白和κ-酪蛋白基因的表达量极显著地高于二维培养(P<0.01)。结果提示,三维培养的BMEC 3种酪蛋白基因的表达量均高于二维培养,三维培养的BMEC在形态上呈现的特点与在体内更加接近。

This study was conducted to investigate the effects of two-and three-dimensional cultivation on cell morphology and gene expressions of β-casein,κ-casein and αs1-casein of bovine mammary epithelial cells(BMECs).Mammary parenchymal tissue was obtained from three healthy Holstein dairy cows aged 5 years.BMECs were purified after cultured for three generations,and then were cultured in two-and three-dimensional system,respectively.Cell morphology and gene expression levels of αs1-casein,β-casein and κ-casein of BMECs cultured in the two systems were compared.The results showed as follows: BMECs cultured in different systems displayed different cell morphologies,and those by two-dimensional cultivation displayed the classical cobblestone morphology,while those by three-dimensional cultivation formed mammospheres-forming aggregates and tubular structures.Gene expressions of αs1-casein(P<0.05),β-casein(P<0.01) and κ-casein(P<0.01) of BMECs by three-dimensional cultivation were significantly higher than those by two-dimensional cultivation.The results indicate that gene expressions of the three caseins of BMECs by three-dimensional cultivation are higher than those by two-dimensional cultivation,and morphological characteristics of three-dimensional cultured BMECs are closely similar to those in vivo.