Bochaton-Piallat M L, Gabbiani F, Ropraz P, Gabbiani G
Department of Pathology, University of Geneva, Switzerland.
Differentiation. 1992 Apr;49(3):175-85. doi: 10.1111/j.1432-0436.1992.tb00665.x.
It is well known that arterial smooth muscle cells (SMC) of adult rats, cultured in a medium containing fetal calf serum (FCS), replicate actively and lose the expression of differentiation markers, such as desmin, smooth muscle (SM) myosin and alpha-SM actin. We report here that compared to freshly isolated cells, primary cultures of SMC from newborn animals show no change in the number of alpha-SM actin containing cells and a less important decrease in the number of desmin and SM myosin containing cells than that seen in primary cultures of SMC from adult animals; moreover, contrary to what is seen in SMC cultured from adult animals, they show an increase of alpha-SM actin mRNA level, alpha-SM actin synthesis and expression per cell. These features are partially maintained at the 5th passage, when the cytoskeletal equipment of adult SMC has further evolved toward dedifferentiation. Cloned newborn rat SMC continue to express alpha-SM actin, desmin and SM myosin at the 5th passage. Thus, newborn SMC maintain, at least in part, the potential to express differentiated features in culture. Heparin has been proposed to control proliferation and differentiation of arterial SMC. When cultured in the presence of heparin, newborn SMC show an increase of alpha-SM actin synthesis and content but no modification of the proportion of alpha-SM actin total (measured by Northern blots) and functional (measured by in vitro translation in a reticulocyte lysate) mRNAs compared to control cells cultured for the same time in FCS containing medium. This suggests that heparin action is exerted at a translational or post-translational level. Cultured newborn rat aortic SMC furnish an in vitro model for the study of several aspects of SMC differentiation and possibly of mechanisms leading to the establishment and prevention of atheromatous plaques.
众所周知,成年大鼠的动脉平滑肌细胞(SMC)在含有胎牛血清(FCS)的培养基中培养时,会积极复制并失去分化标志物的表达,如结蛋白、平滑肌(SM)肌球蛋白和α-SM肌动蛋白。我们在此报告,与新鲜分离的细胞相比,新生动物的SMC原代培养物中,含α-SM肌动蛋白的细胞数量没有变化,含结蛋白和SM肌球蛋白的细胞数量减少程度比成年动物的SMC原代培养物中所见的要小;此外,与成年动物培养的SMC不同,它们显示α-SM肌动蛋白mRNA水平、α-SM肌动蛋白合成以及每个细胞的表达增加。当成年SMC的细胞骨架装备进一步向去分化发展时,这些特征在第5代时仍部分保留。克隆的新生大鼠SMC在第5代时继续表达α-SM肌动蛋白、结蛋白和SM肌球蛋白。因此,新生SMC至少部分保留了在培养中表达分化特征的潜力。肝素已被提议用于控制动脉SMC的增殖和分化。与在含FCS培养基中培养相同时间的对照细胞相比,当在肝素存在下培养时,新生SMC显示α-SM肌动蛋白合成和含量增加,但α-SM肌动蛋白总mRNA(通过Northern印迹法测量)和功能性mRNA(通过网织红细胞裂解物中的体外翻译测量)的比例没有改变。这表明肝素的作用是在翻译或翻译后水平发挥的。培养的新生大鼠主动脉SMC为研究SMC分化的几个方面以及可能导致动脉粥样斑块形成和预防的机制提供了一个体外模型。