The Effect of Blood-Derived Products on the Chondrogenic and Osteogenic Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells Originated from Three Different Locations
The Effect of Blood-Derived Products on the Chondrogenic and Osteogenic Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells Originated from Three Different Locations
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DOI：
10.1155/2019/1358267
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发表时间：
2019-12-31
影响因子：
4.3
通讯作者：
Nehrer, Stefan
中科院分区：
文献类型：
--
作者：
Neubauer, Markus;Kuten, Olga;Nehrer, Stefan
背景。来自脂肪组织的脂肪源性间充质干细胞（AD-MSCs）在关节手术中被认为是“手术废物”，可能为再生医学提供一个强有力的来源。与关节外的非同源脂肪相比，关节内的同源脂肪组织（Hoffa脂肪垫、眼袋脂肪）可能具有更好的成软骨和成骨分化潜力。血液制品可能会进一步增强这种潜力。方法。在全膝关节置换术中，从3个供体的3个部位的脂肪组织中分离出AD-MSCs。用流式细胞术分析分离细胞。细胞补充血液制品：两种富血小板血浆（EDTA存在下制备的EPRP-PRP；柠檬酸盐存在下制备的CPRP-PRP）、超急性血清（hypACT）和标准胎牛血清（FCS）作为阳性对照。采用XTT法检测细胞活力，通过组织学染色和特异性基因表达监测检测细胞分化进展。结果。血液制品促进体外细胞代谢。EDTA-PRP促进软骨形成，柠檬酸PRP促进骨形成，而超急性血清对这两种分化的促进作用比较明显。这一发现在组织学分析和基因表达中是一致的。较低的血制品浓度和较短的分化期导致软骨形成的组织学结果优越。两种类型的PRP有不同的生物效应取决于浓度，而超急性血清似乎有一个更一致的效果，独立于使用的浓度。结论。(i)血液制品制备方法、（ii）抗凝剂类型、（iii）分化时间和（iv）血液制品浓度对干细胞活力和分化潜力有显著影响，有利于不使用抗凝剂、缩短分化时间和降低血液制品浓度。无细胞血液制品，如超急性血清，可以被认为是再生医学，特别是干细胞治疗的替代补充。
Background. Adipose-derived mesenchymal stem cells (AD-MSCs) from fat tissue considered "surgical waste" during joint surgery may provide a potent source for regenerative medicine. Intra-articular, homologous fat tissue (Hoffa's fat pad, pouch fat) might possess a superior chondrogenic and osteogenic differentiation potential in comparison to extra-articular, nonhomologous fat. Blood products might further enhance this potential. Methods. AD-MSCs were isolated from fat tissue of 3 donors from 3 locations each, during total knee replacement. Isolated cells were analyzed via flow cytometry. Cells were supplemented with blood products: two types of platelet-rich plasma (EPRP-PRP prepared in the presence of EDTA; CPRP-PRP prepared in the presence of citrate), hyperacute serum (hypACT), and standard fetal calf serum (FCS) as a positive control. The viability of the cells was determined by XTT assay, and the progress of differentiation was tested via histological staining and monitoring of specific gene expression. Results. Blood products enhance ex vivo cell metabolism. Chondrogenesis is enhanced by EDTA-PRP and osteogenesis by citrate PRP, whereas hyperacute serum enhances both differentiations comparably. This finding was consistent in histological analysis as well as in gene expression. Lower blood product concentrations and shorter differentiation periods lead to superior histological results for chondrogenesis. Both PRP types had a different biological effect depending upon concentration, whereas hyperacute serum seemed to have a more consistent effect, independent of the used concentration. Conclusion. (i) Blood product preparation method, (ii) type of anticoagulant, (iii) differentiation time, and (iv) blood product concentration have a significant influence on stem cell viability and the differentiation potential, favouring no use of anticoagulation, shorter differentiation time, and lower blood product concentrations. Cell-free blood products like hyperacute serum may be considered as an alternative supplementation in regenerative medicine, especially for stem cell therapies.