Kim Eo Jin, Seo Sang Gyo, Shin Hyuk Soo, Lee Doo Jae, Kim Ji Hye, Lee Dong Yeon
Department of Orthopedic Surgery, Hanil General Hospital, Seoul, Korea.
Department of Orthopedic Surgery, Asan Medical Center, Seoul, Korea.
Clin Orthop Surg. 2017 Jun;9(2):239-248. doi: 10.4055/cios.2017.9.2.239. Epub 2017 May 8.
The pericytes in the blood vessel wall have recently been identified to be important in regulating vascular formation, stabilization, remodeling, and function. We isolated and identified pericyte-like platelet-derived growth factor receptor beta-positive (PDGFRβ+) cells from the stromal vascular fraction (SVF) of adipose tissue from critical limb ischemia (CLI) patients and investigated their potential as a reliable source of stem cells for cell-based therapy.
De-identified subcutaneous fat tissues were harvested after amputation in CLI patients. Freshly isolated SVF cells and culture-expanded adipose-derived stem cells (ADSCs) were quantified using flow cytometry. A matrigel tube formation assay and multi-lineage differentiation were performed to assess pericytic and mesenchymal stem cell (MSC)-like characteristics of PDGFRβ+ ADSCs.
PDGFRβ+ cells were located in the pericytic area of various sizes of blood vessels and coexpressed mesenchymal stem cell markers. PDGFRβ+ cells in freshly isolated SVF cells expressed a higher level of stem cell markers (CD34 and CXCR4) and mesenchymal markers (CD13, CD44, CD54, and CD90) than PDGFRβ- cells. expansion of PDGFRβ+ cells resulted in enrichment of the perivascular mesenchymal stem-like (PDGFRβ+/CD90+/CD45-/CD31-) cell fractions. The Matrigel tube formation assay revealed that PDGFRβ+ cells were located in the peritubular area.
PDGFRβ+ ADSCs cells demonstrated a good multilineage differentiation potential. Pericyte-like PDGFRβ+ cells from the SVF of adipose tissue from CLI patients had MSC-like characteristics and could be amplified by culture with preservation of their cell characteristics. We believe PDGFRβ+ cells in the SVF of adipose tissue can be used as a reliable source of stem cells even in CLI patients.
血管壁中的周细胞最近被证实对调节血管形成、稳定、重塑及功能具有重要作用。我们从严重肢体缺血(CLI)患者脂肪组织的基质血管成分(SVF)中分离并鉴定了类周细胞血小板衍生生长因子受体β阳性(PDGFRβ+)细胞,并研究了其作为基于细胞治疗的可靠干细胞来源的潜力。
在CLI患者截肢后采集匿名皮下脂肪组织。使用流式细胞术对新鲜分离的SVF细胞和培养扩增的脂肪来源干细胞(ADSCs)进行定量分析。进行基质胶管形成试验和多向分化实验,以评估PDGFRβ+ ADSCs的周细胞样和间充质干细胞(MSC)样特性。
PDGFRβ+细胞位于不同大小血管的周细胞区域,并共表达间充质干细胞标志物。新鲜分离的SVF细胞中的PDGFRβ+细胞比PDGFRβ-细胞表达更高水平的干细胞标志物(CD34和CXCR4)和间充质标志物(CD13、CD44、CD54和CD90)。PDGFRβ+细胞的扩增导致血管周围间充质干细胞样(PDGFRβ+/CD90+/CD45-/CD31-)细胞组分富集。基质胶管形成试验显示PDGFRβ+细胞位于肾小管周围区域。
PDGFRβ+ ADSCs细胞具有良好的多向分化潜能。CLI患者脂肪组织SVF中的类周细胞PDGFRβ+细胞具有MSC样特性,可通过培养进行扩增并保持其细胞特性。我们认为,即使在CLI患者中,脂肪组织SVF中的PDGFRβ+细胞也可作为可靠的干细胞来源。