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胶原酶自动化处理系统分离间质血管成分的初步研究。

Pilot Study for Isolation of Stromal Vascular Fraction with Collagenase Using an Automated Processing System.

机构信息

Department of Orthopedic Surgery, Hand Unit, The Eisenberg R&D Authority, Shaare Zedek Medical Center, Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem 9103102, Israel.

Department of Plastic & Reconstructive Surgery, Shaare Zedek Medical Center, Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem 9103102, Israel.

出版信息

Int J Mol Sci. 2024 Jun 28;25(13):7148. doi: 10.3390/ijms25137148.

Abstract

There are many potential therapeutic applications for autologous adipose-derived stromal cells. These cells are found in a heterogeneous population isolated from adipose tissue called the stromal vascular fraction (SVF). Closed automated systems are available to release cells from the adherent stroma. Here, we test one system to evaluate the heterogeneous output for yield, purity, cellular characterization, and stemness criteria. The SVF was isolated from three donors using the Automated Cell Station (ACS) from BSL Co., Ltd., Busan, Republic of Korea. The SVF cellular output was characterized for cell yield and viability, immunophenotyping analysis, pluripotent differentiation potential, adhesion to plastic, and colony-forming units. Additionally, the SVF was tested for endotoxin and collagenase residuals. The SVF yield from the ACS system was an average volume of 7.9 ± 0.5 mL containing an average of 19 × 10 nucleated cells with 85 ± 12% viability. Flow cytometry identified a variety of cells, including ASCs (23%), macrophages (24%), endothelial cells (5%), pericytes (4%), and transitional cells (0.5%). The final concentrated product contained cells capable of differentiating into adipogenic, chondrogenic, and osteogenic phenotypes. Furthermore, tests for SVF sterility and purity showed no evidence of endotoxin or collagenase residuals. The ACS system can efficiently process cells from adipose tissue within the timeframe of a single surgical procedure. The cellular characterization indicated that this system can yield a sterile and concentrated SVF output, providing a valuable source of ASCs within the heterogeneous cell population.

摘要

自体脂肪来源的基质细胞有许多潜在的治疗应用。这些细胞存在于从脂肪组织中分离出来的异质群体中,称为基质血管部分(SVF)。封闭的自动化系统可用于从贴壁基质中释放细胞。在这里,我们测试了一种系统,以评估产量、纯度、细胞特征和干性标准的异质输出。SVF 是使用韩国釜山 BSL 有限公司的自动化细胞工作站(ACS)从三个供体中分离出来的。对 SVF 的细胞输出进行了细胞产量和活力、免疫表型分析、多能分化潜能、粘附于塑料和集落形成单位的特性分析。此外,还对 SVF 进行了内毒素和胶原酶残留测试。ACS 系统的 SVF 产量平均为 7.9±0.5mL,含有平均 19×10 个有核细胞,活力为 85±12%。流式细胞术鉴定了多种细胞,包括 ASC(23%)、巨噬细胞(24%)、内皮细胞(5%)、周细胞(4%)和过渡细胞(0.5%)。最终浓缩产物中含有能够分化为成脂、成软骨和成骨表型的细胞。此外,SVF 无菌性和纯度测试表明没有内毒素或胶原酶残留的证据。ACS 系统可以在单次手术过程的时间范围内有效地处理来自脂肪组织的细胞。细胞特征表明,该系统可以产生无菌且浓缩的 SVF 输出,为异质细胞群体中的 ASC 提供了有价值的来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/19a7/11241134/40ac2c45553d/ijms-25-07148-g001.jpg

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