McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA, USA.
J Tissue Eng Regen Med. 2012 Jul;6(7):505-11. doi: 10.1002/term.451. Epub 2011 Jul 14.
Smooth muscle cells (SMCs) are typically used as a cell source for the reconstruction of hollow organs by conventional tissue engineering techniques. However, the necessity for and advantage of the use of tissue-specific SMCs are unknown. The present study investigated the phenotypic changes that occur following isolation and in vitro expansion of rat SMC populations isolated from three different tissues: the aorta, oesophagus and urinary bladder. rSMCs were isolated by enzymatic dispersion and expanded by conventional cell culture techniques, yielding microscopically homogeneous populations. SMC phenotypes were monitored according to their expression of marker proteins during the first two passages. Two of the three SMC populations (rSMC-a and rSMC-e) showed a marked change in their marker protein profiles during the first two passages, which resulted in a homogeneous phenotype that was neither fully contractile nor fully synthetic. SMCs from the urinary bladder did not show such a shift. Differences between the three rSMC populations were observed with regard to proliferative activity and gene expression patterns, suggesting the retention of some tissue-specific cell characteristics. In summary, phenotypic changes in SMCs occur as a result of conventional cell isolation and expansion techniques, thus questioning the necessity for a tissue-specific cell source for regenerative medicine applications.
平滑肌细胞(SMC)通常被用作传统组织工程技术重建中空器官的细胞来源。然而,使用组织特异性 SMC 的必要性和优势尚不清楚。本研究探讨了从三种不同组织(主动脉、食管和膀胱)分离和体外扩增大鼠 SMC 群体后发生的表型变化。通过酶解分散法分离 rSMC,并通过传统细胞培养技术进行扩增,得到了显微镜下均一的细胞群体。在最初的两个传代过程中,根据标记蛋白的表达监测 SMC 表型。三个 SMC 群体中的两个(rSMC-a 和 rSMC-e)在前两个传代过程中标记蛋白谱发生了明显变化,导致形成了一种非完全收缩性也非完全合成性的均一表型。膀胱 SMC 则没有出现这种转变。三种 rSMC 群体之间的差异表现在增殖活性和基因表达模式上,表明保留了一些组织特异性的细胞特征。总之,SMC 的表型变化是由于常规的细胞分离和扩增技术引起的,这使得再生医学应用中对组织特异性细胞来源的必要性受到质疑。