不同保存方法对“临床即用型”脂肪源性干细胞活性影响的评估

摘要

背景：脂肪源性干细胞（Adipose derived stem cells, ADSCs）因其具备来源丰富、取材容易、具有多向分化潜能特点而成为现今成体干细胞研究的热点，同时其在临床研究及应用中也拥有广阔的前景。完善的提取设施及最符合临床使用规范的细胞使用及保存方法，是ADSCs能更安全有效的应用于临床的关键所在。目的：通过对现有的细胞保存介质、保存方法及保存时间进行综述，为临床即用型脂肪源性干细胞的短期运送及保存提供理论基础。方法：广泛查阅近年相关文献，对脂肪源性干细胞的研究历史、临床应用及其他成体干细胞的保存方式及临床转化应用进行综合论述。结果：4℃条件下保存细胞，对短期内细胞的活性和生物学特性影响最小，相对室温条件下有明显优势；血清是细胞培养中重要的成分之一，包含各种血浆蛋白、生长因子、碳水化合物等多种物质，能提供氨基酸、维生素、无机物等细胞生长必需物质，对脂肪源性干细胞的存活力及细胞增殖亦有促进作用；富血小板血浆（Platelet-rich plasma,PRP）内含丰富的血小板，是全血中血小板数量3倍以上，能释放出多种生长因子，如血小板衍生生长因子（Platelet derived growth factor,PDGF）、表皮生长因子（Epidermal growth factor,EGF）、类胰岛素生长因子（Insulin like growth factor,IGF）等，对脂肪源性干细胞的细胞增殖及分化功能有明显的促进作用。结论：4℃条件下使用含丰富的生长因子及无毒性的血清及PRP可能是较好的保存细胞活性及能安全使用的保存介质。%Background Adipose derived stem cells as immediate areas of research focus, because it is plentiful, easily separated, multi-directional differentiation potential, secretion and other biological characteristics, it has broad prospect in clinical research and application. Perfect facilities and good cells storage method can better assurance the safety and efficacy of cell therapy. Objective Through the review of existing cell preservation media and the storage methods and the storage time to provide a theoretical basis for the clinical ready-to-use adipose derived stem cells in transit or in the short term of related research. Methods Review of related literature in recent years, discuss comprehensively from ADSCs history,clinical application and preservation way of the other adult stem cells and so on. Results The stem cells stored at 4℃ better than at home temperature,and no effect on the biological characteristics of cells in short term. Serum is important for ADSCs, it is contain plasma proteins、growth factor、carbohydrate and so on,with cell growth elements, such as amino acids,vitamins ,etc. Platelet in platelet rich plasma (PRP) is three times more than whole blood platelets can release a lot of growth factors, such as PDGF,EGF,IGF, etc. On the cellula proliferation and differentiation ability of adipose derived stem cells have obvious role in promoting. Conclusion In order to maintain the best cell activity effectively, adipose derived stem cells in the short-term preservation or transit, cells had better preserved in a solution containing PRP or serum at 4℃.