Comparison of clinical grade human platelet lysates for cultivation of mesenchymal stromal cells from bone marrow and adipose tissue
Comparison of clinical grade human platelet lysates for cultivation of mesenchymal stromal cells from bone marrow and adipose tissue
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DOI：
10.3109/00365513.2015.1099723
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发表时间：
2016-02-17
影响因子：
2.1
通讯作者：
Haack-Sorensen, Mandana
中科院分区：
文献类型：
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作者：
Juhl, Morten;Tratwal, Josefine;Haack-Sorensen, Mandana
背景：间充质基质细胞（MSCs）在再生医学治疗应用中的效用已受到广泛关注。基于MSC的方法的临床转化需要体外培养扩增以获得足够数量的细胞。理想的细胞培养基应不含任何动物源性成分。我们已经评估了人类血小板裂解物（hPL）是否可以成为动物补充剂的有吸引力的替代品。研究方法：将从三个供体获得的来自骨髓的MSC（BMSC）和脂肪组织来源的基质细胞（ASC）在满足临床使用的良好制造规范标准的三种不同的市售hPL中培养扩增。将在补充有5% PLT-Max（米尔克里克）、Stemulate（TM）PL-S和Stemulate（TM）PL-SP（COOK General Biotechnology）的最低必需培养基α中培养的BMSC和ASC与具有10%胎牛血清（FBS）的标准培养条件进行比较。分析细胞形态、增殖、表型、基因组稳定性和分化潜能。结果如下：与制造商无关，与FBS培养基相比，在hPL培养基中培养的BMSC和ASC显示出增殖能力的显著增加。一般来说，在hPL培养基扩增后，BMSC和ASC的免疫表型均符合国际细胞治疗学会（ISCT）标准。比较基因组杂交测量表明，在hPL培养基或FBS培养基中培养的BMSC或ASC没有不平衡的染色体重排。在所有四种培养条件下，BMSCs和ASCs均可分化为成骨、成脂或成软骨谱系。结论：所有三种临床批准的商业人血小板裂解物加速BMSC和ASC的增殖，并且细胞满足ISCT间充质表型要求而不表现出染色体畸变。
Background: The utility of mesenchymal stromal cells (MSCs) in therapeutic applications for regenerative medicine has gained much attention. Clinical translation of MSC-based approaches requires in vitro culture-expansion to achieve a sufficient number of cells. The ideal cell culture medium should be devoid of any animal derived components. We have evaluated whether human Platelet Lysate (hPL) could be an attractive alternative to animal supplements. Methods: MSCs from bone marrow (BMSCs) and adipose tissue-derived stromal cells (ASCs) obtained from three donors were culture expanded in three different commercially available hPL fulfilling good manufacturing practice criteria for clinical use. BMSCs and ASCs cultured in Minimum Essential Medium Eagle-alpha supplemented with 5% PLT-Max (Mill Creek), Stemulate (TM) PL-S and Stemulate (TM) PL-SP (COOK General Biotechnology) were compared to standard culture conditions with 10% fetal bovine serum (FBS). Cell morphology, proliferation, phenotype, genomic stability, and differentiation potential were analyzed. Results: Regardless of manufacturer, BMSCs and ASCs cultured in hPL media showed a significant increase in proliferation capacity compared to FBS medium. In general, the immunophenotype of both BMSCs and ASCs fulfilled International Society for Cellular Therapy (ISCT) criteria after hPL media expansion. Comparative genomic hybridization measurements demonstrated no unbalanced chromosomal rearrangements for BMSCs or ASCs cultured in hPL media or FBS medium. The BMSCs and ASCs could differentiate into osteogenic, adipogenic, or chondrogenic lineages in all four culture conditions. Conclusion: All three clinically approved commercial human platelet lysates accelerated proliferation of BMSCs and ASCs and the cells meet the ISCT mesenchymal phenotypic requirements without exhibiting chromosomal aberrations.