胎牛血清(FBS),是体外培养最常用的培养基添加剂,尽管它的组成不确定,其潜在的免疫原性和可能的朊病毒/人畜共患传播。由于这些原因,已经做出了大量努力来寻找替代品,例如无血清培养基或人血小板裂解物(hPL)。我们的目标是在已发表的文献中批判性地评估hPL的最新技术,将其影响与FBS进行比较。
2019年6月,对整个WebofScience进行了系统搜索,Medline和PubMed数据库用以下搜索项进行:(间充质干细胞)AND(胎牛血清或胎牛小牛)AND(人血小板裂解物)。从这次搜索中排除了2005年之前发表的评论文章,其中间充质干细胞(MSC)不是来自人类来源的手稿,以及FBS控件丢失时。
基于我们的搜索算法,共入选56篇论文。对这些论文的综述表明,与hPL一起培养的hMSCs表现出纺锤形的细长形态,有较高的增殖指数,相似的分化(CD)标记簇,分化谱系没有显著变化(骨细胞,脂肪细胞,和软骨细胞)与用FBS培养的细胞相比。原代hMSC的主要来源是脂肪组织或骨髓;在一些研究中,从替代来源分离的细胞在它们的应答中没有显示相关差异。
尽管介质选择不同,但hPL制造缺乏标准化,大多数出版物支持hPL在促进粘附方面至少与FBS一样有效,hMSCs的存活和增殖。我们得出的结论是,在hMSCs培养中,hPL应被视为FBS的可行替代品,尤其是在临床应用方面。
Foetal bovine serum (FBS), is the most commonly used culture medium additive for in vitro cultures, despite its undefined composition, its potential immunogenicity and possible prion/zoonotic transmission. For these reasons, significant efforts have been targeted at finding a substitute, such as serum free-media or human platelet-lysates (hPL). Our aim is to critically appraise the state-of-art for hPL in the published literature, comparing its impact with FBS.
In June 2019 a systematic search of the entire Web of Science, Medline and PubMed database was performed with the following search terms: (mesenchymal stem cells) AND (fetal bovine serum OR fetal bovine calf) AND (human platelet lysate). Excluded from this search were
review articles that were published before 2005, manuscripts in which mesenchymal stem cells (MSCs) were not from human sources, and when the FBS controls were missing.
Based on our search algorithm, 56 papers were selected. A
review of these papers indicated that hMSCs cultured with hPL showed a spindle-shaped elongated morphology, had higher proliferation indexes, similar cluster of differentiation (CD) markers and no significant variation in differentiation lineage (osteocyte, adipocyte, and chondrocyte) compared to those cultured with FBS. Main sources of primary hMSCs were either fat tissue or bone marrow; in a few studies cells isolated from alternative sources showed no relevant difference in their response.
Despite the difference in medium choice and a lack of standardization of hPL manufacturing, the majority of publications support that hPL was at least as effective as FBS in promoting adhesion, survival and proliferation of hMSCs. We conclude that hPL should be considered a viable alternative to FBS in hMSCs culture-especially with a view for their clinical use.