Low serum and serum-free culture of multipotential human adipose stem cells
Low serum and serum-free culture of multipotential human adipose stem cells
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DOI：
10.1080/14653240701508452
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发表时间：
2007-01-01
期刊：
影响因子：
4.5
通讯作者：
Katz, A. J.
中科院分区：
文献类型：
--
作者：
Parker, A. M.;Shang, H.;Katz, A. J.
研究背景脂肪组织为转化性临床研究提供了一种简单、易得和丰富的干细胞来源。目前流行的培养技术包括使用FBS，这是一种高度可变和未定义的成分，可能会带来患者的不良反应。为了避免在人类脂肪干细胞(ASC)培养中使用动物产品，我们开发了两种新的培养方法，一种是非常低的人血清扩增液，另一种是完全无血清的培养液。方法通过系列试验，我们开发了一种高度浓缩的培养液配方，可以在有或没有添加0.5%人血清的情况下使用，这一比例很容易从自体血液中获得。结果在非常低的血清培养条件下，早期传代的细胞群体倍增时间平均为1.86天，而无血清配方的细胞生长较差，倍增时间平均为5.79天。在这两种条件下的ASC在体外都保持了分化为脂肪、软骨和成骨细胞的能力，尽管早期传代时CD34的表达较低。ALDH、HLA、CD133、CD184和CD31的表达与在10%FBS中培养的细胞相似。讨论这些新的培养条件提供了一个独特的环境，可以在不受动物血清干扰的情况下研究ASCs，并允许在无动物产品的环境中快速扩增自体ASCs，用于人类临床试验。
BackgroundAdipose tissue provides an easily, accessible and abundant source of putative stem cells for translational clinical research. Currently prevalent culture techniques include the use of FBS, a highly variable and undefined component, which brings with it the potential for adverse patient reactions. In an effort to eliminate the use of animal products in human adipose stem cell (ASC) cultures, we have developed two new culture methods, a very low human serum expansion medium and a completely serum-free medium.MethodsThrough serial testing, a highly enriched medium formulation was developed for use with and without the addition of 0.5% human serum, an amount easily obtainable from autologous blood draws.ResultsVery low-serum culture yielded population-doubling times averaging 1.86 days in early passage, while the serum-free formulation was associated with less robust cell growth, with doubling times averaging 5.79 days. ASC in both conditions maintained its ability to differentiate into adipo-, chondro- and osteogenic lineages in vitro, despite lower expression of CD34 in early passage. Expression of ALDH, HLA, CD133, CD184, and CD31 was comparable with that seen in cells cultured in 10% FBS.DiscussionThese newly developed culture conditions provide a unique environment within which to study ASCs without the interference of animal serum, and allow for the rapid expansion of autologous ASCs in culture in an animal product-free environment for use in human clinical trials.