Heydarkhan-Hagvall Sepideh, Schenke-Layland Katja, Yang Jin Q, Heydarkhan Sanaz, Xu Yuhuan, Zuk Patricia A, MacLellan W Robb, Beygui Ramin E
Regenerative Bioengineering and Repair Laboratory, Department of Surgery, David Geffen School of Medicine at UCLA, Los Angeles, Calif., USA.
Cells Tissues Organs. 2008;187(4):263-74. doi: 10.1159/000113407. Epub 2008 Jan 14.
BACKGROUND/AIMS: A crucial step in providing clinically relevant applications of cardiovascular tissue engineering involves the identification of a suitable cell source. The objective of this study was to identify the exogenous and endogenous parameters that are critical for the differentiation of human adipose stem cells (hASCs) into cardiovascular cells.
hASCs were isolated from human lipoaspirate samples, analyzed, and subjected to two differentiation protocols.
As shown by fluorescence-activated cell sorter (FACS) analysis, a population of hASCs expressed stem cell markers including CXCR4, CD34, c-kit, and ABCG2. Further, FACS and immunofluorescence analysis of hASCs, cultured for 2 weeks in DMEM-20%-FBS, showed the expression of smooth muscle cell (SMC)-specific markers including SM alpha-actin, basic calponin, h-caldesmon and SM myosin. hASCs, cultured for 2 weeks in endothelial cell growth medium-2 (EGM-2), formed a network of branched tube-like structures positive for CD31, CD144, and von Willebrand factor. The frequency of endothelial cell (EC) marker-expressing cells was passage number-dependent. Moreover, hASCs attached and formed a confluent layer on top of electrospun collagen-elastin scaffolds. Scanning electron microscopy and DAPI staining confirmed the integration of hASCs with the fibers and formation of a cell-matrix network.
Our results indicate that hASCs are a potential cell source for cardiovascular tissue engineering; however, the differentiation capacity of hASCs into SMCs and ECs is passage number- and culture condition-dependent.
背景/目的：在心血管组织工程临床相关应用中，关键的一步是确定合适的细胞来源。本研究的目的是确定对人脂肪干细胞（hASC）分化为心血管细胞至关重要的外源性和内源性参数。
从人抽脂样本中分离、分析hASC，并使其经历两种分化方案。
荧光激活细胞分选仪（FACS）分析显示，一群hASC表达包括CXCR4、CD34、c-kit和ABCG2在内的干细胞标志物。此外，在含20%胎牛血清的DMEM中培养2周的hASC的FACS和免疫荧光分析显示，平滑肌细胞（SMC）特异性标志物包括平滑肌α-肌动蛋白、碱性钙调蛋白、h-钙调蛋白和平滑肌肌球蛋白的表达。在内皮细胞生长培养基-2（EGM-2）中培养2周的hASC形成了对CD31、CD144和血管性血友病因子呈阳性的分支管状结构网络。表达内皮细胞（EC）标志物的细胞频率与传代次数有关。此外，hASC附着在电纺胶原-弹性蛋白支架上并形成汇合层。扫描电子显微镜和DAPI染色证实了hASC与纤维的整合以及细胞-基质网络的形成。
我们的结果表明，hASC是心血管组织工程的潜在细胞来源；然而，hASC向SMC和EC的分化能力取决于传代次数和培养条件。
