Department of Hematology-Oncology, Wonju College of Medicine, Yonsei University, Wonju, Korea.
Yonsei Med J. 2011 Nov;52(6):999-1007. doi: 10.3349/ymj.2011.52.6.999.
This study examined a rapid isolation method decreasing the time and cost of the clinical application of adipose tissue-derived stem cells (ASCs).
Aliquots (10 g) of the lipoaspirates were stored at 4°C without supplying oxygen or nutrients. At the indicated time points, the yield of mononuclear cells was evaluated and the stem cell population was counted by colony forming unit-fibroblast assays. Cell surface markers, stem cell-related transcription factors, and differentiation potentials of ASCs were analyzed.
When the lipoaspirates were stored at 4°C, the total yield of mononuclear cells decreased, but the stem cell population was enriched. These ASCs expressed CD44, CD73, CD90, CD105, and HLA-ABC but not CD14, CD31, CD34, CD45, CD117, CD133, and HLA-DR. The number of ASCs increased 1×10(14) fold for 120 days. ASCs differentiated into osteoblasts, adipocytes, muscle cells, or neuronal cells.
ASCs isolated from lipoaspirates and stored for 24 hours at 4°C have similar properties to ASCs isolated from fresh lipoaspirates. Our results suggest that ASCs can be isolated with high frequency by optimal storage at 4°C for 24 hours, and those ASCs are highly proliferative and multipotent, similar to ASCs isolated from fresh lipoaspirates. These ASCs can be useful for clinical application because they are time- and cost-efficient, and these cells maintain their stemness for a long time, like ASCs isolated from fresh lipoaspirates.
本研究旨在探索一种快速分离方法，以减少脂肪组织源性干细胞（ADSCs）临床应用的时间和成本。
将 10 g 份的脂肪抽吸物储存在 4°C 下，不提供氧气或营养物质。在指定的时间点，评估单核细胞的产量，并通过集落形成单位-成纤维细胞测定法计数干细胞群体。分析 ADSC 的细胞表面标志物、干细胞相关转录因子和分化潜能。
当脂肪抽吸物储存在 4°C 时，单核细胞的总产量减少，但干细胞群体得到了富集。这些 ADSC 表达 CD44、CD73、CD90、CD105 和 HLA-ABC，但不表达 CD14、CD31、CD34、CD45、CD117、CD133 和 HLA-DR。在 120 天内，ADSC 的数量增加了 1×10(14)倍。ADSC 可分化为成骨细胞、脂肪细胞、肌肉细胞或神经元细胞。
从脂肪抽吸物中分离并在 4°C 下储存 24 小时的 ADSC 具有与从新鲜脂肪抽吸物中分离的 ADSC 相似的特性。我们的结果表明，通过在 4°C 下最佳储存 24 小时，可以以较高的频率分离 ADSC，这些 ADSC 具有较高的增殖能力和多能性，与从新鲜脂肪抽吸物中分离的 ADSC 相似。这些 ADSC 可用于临床应用，因为它们省时、省钱，并且这些细胞长时间保持其干细胞特性，就像从新鲜脂肪抽吸物中分离的 ADSC 一样。
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