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从髌下脂肪垫中分离出来的新鲜基质细胞适合用于一步手术来再生软骨组织。

Freshly isolated stromal cells from the infrapatellar fat pad are suitable for a one-step surgical procedure to regenerate cartilage tissue.

机构信息

Department of Plastic, Reconstructive and Hand Surgery, VU University Medical Center, Amsterdam, the Netherlands.

出版信息

Cytotherapy. 2009;11(8):1052-64. doi: 10.3109/14653240903219122.

Abstract

BACKGROUND AIMS

Stem cell therapies are being evaluated as promising alternatives for cartilage regeneration. We investigated whether stromal vascular fraction cells (SVF) from the infrapatellar (Hoffa) fat pad are suitable for a one-step surgical procedure to treat focal cartilage defects.

METHODS

SVF was harvested from patients undergoing knee arthroplasty (n = 53). Colony-forming unit (CFU) assays, growth kinetics and surface marker profiles were determined, and the chondrogenic differentiation capacity of freshly isolated SVF was assessed after seeding in three-dimensional poly (L-lactic-co-epsilon-caprolactone) scaffolds.

RESULTS

SVF yield per fat pad varied between 0.55 and 16 x 10(6) cells. CFU frequency and population doubling time were 2.6 +/- 0.6% and +/-2 days, respectively. Surface marker profiles matched those of subcutaneous-derived adipose-derived stem cells (ASC). CFU from Hoffa SVF showed differentiation toward osteogenic and adipogenic lineages. Cartilage differentiation was confirmed by up-regulation of the cartilage genes sox9, aggrecan, collagen type II and cartilage oligomeric matrix protein (COMP), collagen II immunostaining, Alcian Blue staining and glycosaminoglycan production. Compared with passaged cells, SVF showed at least similar chondrogenic potential.

CONCLUSIONS

This study demonstrates that SVF cells from the infrapatellar fat pad are suitable for future application in a one-step surgical procedure to regenerate cartilage tissue. SVF shows similar favorable characteristics as cultured ASC, and chondrogenic differentiation even appears to be slightly better. However, because of variable harvesting volumes and yields, SVF from the infrapatellar fat pad might only be applicable for treatment of small focal cartilage defects, whereas for larger osteoarthritic defects subcutaneous adipose tissue depot would be preferable.

摘要

背景目的

干细胞疗法被评估为软骨再生的有前途的替代方法。我们研究了髌下(Hoffa)脂肪垫的基质血管成分(SVF)是否适合用于一步手术来治疗局灶性软骨缺损。

方法

从接受膝关节置换术的患者中采集 SVF(n = 53)。测定集落形成单位(CFU)测定、生长动力学和表面标志物谱,并在三维聚(L-丙交酯-co-ε-己内酯)支架上接种后评估新鲜分离的 SVF 的软骨分化能力。

结果

每个脂肪垫的 SVF 产量在 0.55 至 16 x 10(6)个细胞之间变化。CFU 频率和倍增时间分别为 2.6 +/- 0.6%和 +/-2 天。表面标志物谱与皮下脂肪衍生的脂肪干细胞(ASC)相匹配。Hoffa SVF 的 CFU 显示向成骨和脂肪谱系分化。软骨分化通过软骨基因 Sox9、聚集蛋白聚糖、II 型胶原和软骨寡聚基质蛋白(COMP)的上调、胶原 II 免疫染色、阿尔辛蓝染色和糖胺聚糖产生得到证实。与传代细胞相比,SVF 显示出至少相似的软骨生成潜力。

结论

本研究表明,髌下脂肪垫的 SVF 细胞适合用于未来的一步手术程序来再生软骨组织。SVF 显示出与培养的 ASC 相似的有利特征,并且软骨分化似乎甚至更好。然而,由于收获体积和产量的变化,髌下脂肪垫的 SVF 可能仅适用于治疗小的局灶性软骨缺损,而对于较大的骨关节炎缺损,皮下脂肪组织库将是首选。

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