Differentiation of smooth muscle progenitor cells in peripheral blood and its application in tissue engineered blood vessels

查看全文 作　　者：Shang-zhe [1]XIE;Ning-tao [1]FANG;Shui [1]LIU;Ping [2]ZHOU;Yi [2]ZHANG;Song-mei [1]WANG;Hong-yang [1]GAO;Luan-feng [1]PAN

机构地区：[1]Laboratory of Molecular Biology, Shanghai Medical College, Fudan University, Shanghai 200032, China;[2]Key Laboratory of Molecular Engineering of Polymers, Ministry of Education, Department of Macromolecular Science, Fudan University, Shanghai 200433, China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》索引2008年第9卷第12期,共8页

基　　金：supported by Shanghai Science Committee Fund for Key Research Project (No. 04JC14012);Fudan University Med-X Fund, China 摘　　要：Background: A major shortcoming in tissue engineered blood vessels (TEBVs) is the lack of healthy and easily attainable smooth muscle cells (SMCs). Smooth muscle progenitor cells (SPCs), especially from peripheral blood, may offer an alternative cell source for tissue engineering involving a less invasive harvesting technique. Methods: SPCs were isolated from 5-ml fresh rat peripheral blood by density-gradient centrifugation and cultured for 3 weeks in endothelial growth medium-2-MV (EGM-2-MV) medium containing platelet-derived growth factor-BB (PDGF BB). Before seeded on the synthesized scaffold, SPC-derived smooth muscle outgrowth cell (SOC) phenotypes were assessed by immuno-fluorescent staining, Western blot analysis, and reverse transcription polymerase chain reaction (RT-PCR). The cells were seeded onto the silk fibroin-modified poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (SF-PHBHHx) scaffolds by 6×104 cells/cm2 and cultured under the static con- dition for 3 weeks. The growth and proliferation of the seeded cells on the scaffold were analyzed by 3-(4,5-dimethylthiazol-2-yl)- diphenyltetrazolium bromide (MTT) assay, scanning electron microscope (SEM), and 4,6-diamidino-2-phenylindole (DAPI) staining. Results: SOCs displayed specific 'hill and valley' morphology, expressed the specific markers of the SMC lineage: smooth muscle (SM) α-actin, calponin and smooth muscle myosin heavy chain (SM MHC) at protein and messenger ribonucleic acid (mRNA) levels. RT-PCR results demonstrate that SOCs also expressed smooth muscle protein 22α (SM22α), a contractile protein, and extracellular matrix components elastin and matrix Gla protein (MGP), as well as vascular endothelial growth factor (VEGF). After seeded on the SF-PHBHHx scaffold, the cells showed excellent metabolic activity and proliferation. Conclusion: SPCs isolated from peripheral blood can be differentiated into the SMCs in vitro and have an impressive growth potential in the biodegradable synthesized scaffold. Thus, SPCs may be a promising cell source for constructing TEBVs. 关键词：细胞分子生物学单元细胞工程师经验

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Differentiation of smooth muscle progenitor cells in peripheral blood and its application in tissue engineered blood vessels

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