从脂肪组织源性基质血管部分离的最佳条件，用于开发自动化系统。
Optimal Condition of Isolation from an Adipose Tissue-Derived Stromal Vascular Fraction for the Development of Automated Systems.
机构信息
Department of Biomedicine and Health Sciences, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul, 06591, Republic of Korea.
Department of Plastic and Reconstructive Surgery, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul, 06591, Republic of Korea.
出版信息
Tissue Eng Regen Med. 2020 Apr;17(2):203-208. doi: 10.1007/s13770-019-00238-3. Epub 2020 Jan 29.
BACKGROUND
The stromal vascular fraction (SVF) isolated from adipose tissue, which contains stem cells as well as other cell types, has been applied in various research fields. Although different enzymatic concentrations and treatment durations have been applied to isolate the SVF, optimal conditions have not been established. Thus, we aimed to establish the optimal conditions for isolation of the SVF from adipose tissue by automated systems.
METHODS
The SVF was collected from removed adipose tissues of five donors during surgery. The SVF was treated with 0.1% or 0.2% collagenase type I for 20, 40, or 60 min. Then, colony forming unit (CFU) assays and flow cytometry were performed to characterize the adipose stem cells (ASCs). A cytokine array was used to investigate the correlation between colony-formation ability and the secretion of isolated ASCs.
RESULTS
Treatment with 0.1% collagenase type I for 60 min resulted in a higher SVF yield, whereas treatment with 0.1% collagenase for 40 min resulted in higher CFU values. In addition, expression of interleukin (IL)-6, IL-8, and monocyte chemoattractant protein-1 in the SVF was higher in the high-CFU group than in the low-CFU group.
CONCLUSION
The optimal conditions for isolation of the SVF from adipose tissue were treatment with 0.1% collagenase type I for 40 min. We identified the conditions required for efficient SVF isolation based on high CFU values, and our results will facilitate the development of automated systems.
背景
从脂肪组织中分离出来的基质血管部分（SVF）含有干细胞和其他细胞类型，已应用于各个研究领域。虽然已经应用了不同的酶浓度和处理时间来分离 SVF，但尚未建立最佳条件。因此，我们旨在通过自动化系统确定从脂肪组织中分离 SVF 的最佳条件。
方法
从手术过程中移除的 5 位供体的脂肪组织中收集 SVF。用 0.1%或 0.2%的Ⅰ型胶原酶处理 20、40 或 60 分钟。然后进行集落形成单位（CFU）测定和流式细胞术，以表征脂肪干细胞（ASC）。细胞因子阵列用于研究集落形成能力与分离的 ASC 分泌之间的相关性。
结果
用 0.1%的Ⅰ型胶原酶处理 60 分钟可获得更高的 SVF 产量，而用 0.1%的胶原酶处理 40 分钟可获得更高的 CFU 值。此外，在高 CFU 组中，SVF 中白细胞介素（IL）-6、IL-8 和单核细胞趋化蛋白-1 的表达高于低 CFU 组。
结论
从脂肪组织中分离 SVF 的最佳条件是用 0.1%的Ⅰ型胶原酶处理 40 分钟。我们根据高 CFU 值确定了分离高效 SVF 所需的条件，我们的结果将有助于自动化系统的发展。
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