Fujita H, Shimokado K, Yutani C, Takaichi S, Masuda J, Ogata J
Department of Pathology, National Cardiovascular Center, Osaka, Japan.
Exp Mol Pathol. 1993 Feb;58(1):25-39. doi: 10.1006/exmp.1993.1003.
Neonatal vascular smooth muscle cells (SMC) in culture have been demonstrated to be quite different from adult SMC and to be similar to intimal SMC in animal models. To characterize human neonatal vascular SMC in culture, cultures of arterial SMC were prepared by an explant method from the subclavian arteries of autopsied patients (10 adults and 6 neonates). The morphology and growth characteristics of these cells were compared. All cells were positively immunostained with HHF 35, a monoclonal antibody specific for muscle actin. Electron microscopically, both adult and neonatal SMC were of synthetic phenotype. SMC from neonates had a short population doubling time (PDT, 28.6 +/- 7.5 hr) and high saturation density (SD, 37.5 +/- 11.9 x 10(4) cells/cm2). They did not show hill and valley growth patterns. Among the SMC cultured from adult media, two subtypes were distinguished, based on their growth characteristics. Classical adult SMC (7 of 10 cases) grew in hill and valley patterns with long PDT and low SD values (61.7 +/- 28.8 hr, 6.5 +/- 1.9 x 10(4) cells/cm2, respectively). The second subtype (3 of 10 cases), neonatal-type adult SMC, had PDT and SD values (22.9 +/- 4.0 hr, 31.3 +/- 14.7 x 10(4) cells/cm2, respectively) similar to those of neonatal SMC. Intimal SMC became senescent in early phases of subculture. To test for the possible participation of autocrine growth factors in the heterogeneity of the growth patterns, Northern blot analysis was conducted for PDGF-A, TGF-beta, c-myc, and c-fos mRNA in three types of SMC. There was no significant difference in these mRNA levels between the SMC. We demonstrated that human neonatal vascular SMC in culture are quite different in their growth characteristics from classical adult SMC in culture and that neonatal-type SMC can be isolated from adult media.
培养的新生儿血管平滑肌细胞（SMC）已被证明与成人SMC有很大不同，且与动物模型中的内膜SMC相似。为了表征培养的人新生儿血管SMC，通过外植法从尸检患者（10名成人和6名新生儿）的锁骨下动脉制备动脉SMC培养物。比较了这些细胞的形态和生长特性。所有细胞均用HHF 35进行阳性免疫染色，HHF 35是一种对肌肉肌动蛋白特异的单克隆抗体。电子显微镜下，成人和新生儿SMC均为合成表型。新生儿的SMC群体倍增时间短（PDT，28.6±7.5小时），饱和密度高（SD，37.5±11.9×10⁴个细胞/cm²）。它们没有显示出山丘和山谷生长模式。在从成人中膜培养的SMC中，根据其生长特性区分出两种亚型。典型的成人SMC（10例中的7例）以山丘和山谷模式生长，PDT长，SD值低（分别为61.7±28.8小时，6.5±1.9×10⁴个细胞/cm²）。第二种亚型（10例中的3例），即新生儿型成人SMC，其PDT和SD值（分别为22.9±4.0小时，31.3±14.7×10⁴个细胞/cm²）与新生儿SMC相似。内膜SMC在传代培养的早期阶段就衰老了。为了测试自分泌生长因子是否可能参与生长模式的异质性，对三种类型的SMC中的血小板衍生生长因子-A（PDGF-A）、转化生长因子-β（TGF-β）、c-myc和c-fos mRNA进行了Northern印迹分析。这些SMC之间的这些mRNA水平没有显著差异。我们证明，培养的人新生儿血管SMC在生长特性上与培养的典型成人SMC有很大不同，并且可以从成人中膜中分离出新生儿型SMC。
