The Infrapatellar Fat Pad as a Source of Perivascular Stem Cells with Increased Chondrogenic Potential for Regenerative Medicine
The Infrapatellar Fat Pad as a Source of Perivascular Stem Cells with Increased Chondrogenic Potential for Regenerative Medicine
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DOI：
10.5966/sctm.2016-0040
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发表时间：
2017-01-01
影响因子：
6
通讯作者：
Peault, Bruno
中科院分区：
文献类型：
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作者：
Hindle, Paul;Khan, Nusrat;Peault, Bruno
血管周围干细胞(PSCs)是间充质干细胞(MSCs)的天然祖先，是体内负责内环境稳定和修复的干细胞。前瞻性地鉴定和分离的PSCs表现出更强的可塑性和成骨潜能。与来自皮下脂肪的细胞相比，来自髌下脂肪垫(IFP)的细胞显示出更强的软骨形成潜力。这项研究评估了IFP PSCs与来自IFP和骨髓的MSCs的软骨形成潜力。免疫组织化学显示血管周围标志物(CD146、CD34、神经/胶质抗原2[NG2]、血小板衍生生长因子受体β[PDGFRβ]和α-平滑肌肌动蛋白[α-SMA])与内皮标志物(CD31、CD144、von Willebrand因子[vWF])的定位有关。用流式细胞术从间质血管部分(分别为3.8%和21.2%)分离出周细胞和外膜细胞，存活率为88%。平均分离的周细胞数和外膜细胞数分别为4.6+/-2.2×10(4)和16.2+/-3.2×10(4)，相当于每克组织分离出7.96+/-4.4×10(3)和20.8+/-4.3×10(3)个细胞。荧光激活细胞分选显示培养的PSCs为CD44+CD90+CD105+；聚合酶链式反应和免疫细胞化学显示周细胞保持CD146+表型，并表达周细胞标志物PDGFRβ和NG2。组织化学染色和基因表达证实细胞分化。在颗粒模型中，IFP PSCs和MSCs产生的细胞外基质明显多于骨髓MSCs(p
Perivascular stem cells (PSCs) are the natural ancestors of mesenchymal stem cells (MSCs) and are the stem cells responsible for homeostasis and repair in vivo. Prospectively identified and isolated PSCs have demonstrated increased plasticity and osteogenic potential. Cells from the infrapatellar fat pad (IFP) have demonstrated increased chondrogenic potential compared with those from subcutaneous fat. This research assessed the chondrogenic potential of IFP PSCs compared with MSCs from the IFP and bone marrow. Immunohistochemistry demonstrated the location of perivascular markers (CD146, CD34, neural/glial antigen 2 [NG2], platelet-derived growth factor receptor-beta [PDGFR beta], and alpha-smooth muscle actin [alpha-SMA]) in relation to endothelial markers (CD31, CD144, von Willebrand factor [vWF]). Pericytes and adventitial cells were isolated from the stromal vascular fraction (3.8% and 21.2%, respectively) using flow cytometry with a viability of 88%. The mean numbers of pericytes and adventitial cells isolatedwere 4.6 +/- 2.2 x 10(4) and 16.2 +/- 3.2 x 10(4), respectively, equating to 7.96 +/- 4.4 x 10(3) and 20.8 +/- 4.3 x 10(3) cells per gram of harvested tissue. Fluorescence-activated cell sorting demonstrated that cultured PSCs were CD44+CD90+CD105+; polymerase chain reaction and immunocytochemistry demonstrated that pericytes retained their CD146+ phenotype and expressed the pericyte markers PDGFR beta and NG2. Differentiation was confirmed using histochemical stains and genetic expression. Using a pellet model, the IFP PSCs and the MSCs generated significantly more extracellular matrix than bone marrow MSCs (p