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化学定义的无动物来源和无血清细胞培养液，用于培养人脂肪干细胞。

Chemically Defined Xeno- and Serum-Free Cell Culture Medium to Grow Human Adipose Stem Cells.

机构信息

Foundation for Cardiological Research and Education (FCRE), Cardiocentro Ticino Foundation, 6807 Taverne, Switzerland.

Institute of Chemistry & Biotechnology, Competence Center of Biochemical Engineering & Cell Cultivation Technique Zurich University of Applied Sciences, 8820 Wädenswil, Switzerland.

出版信息

Cells. 2021 Feb 22;10(2):466. doi: 10.3390/cells10020466.

DOI:10.3390/cells10020466
PMID:33671568
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7926673/
Abstract

Adipose tissue is an abundant source of stem cells. However, liposuction cannot yield cell quantities sufficient for direct applications in regenerative medicine. Therefore, the development of GMP-compliant ex vivo expansion protocols is required to ensure the production of a "cell drug" that is safe, reproducible, and cost-effective. Thus, we developed our own basal defined xeno- and serum-free cell culture medium (), specifically formulated to grow human adipose stem cells (hASCs). With this medium, we can directly culture the stromal vascular fraction (SVF) cells in defined cell culture conditions to obtain hASCs. Cells proliferate while remaining undifferentiated, as shown by Flow Cytometry (FACS), Quantitative Reverse Transcription PCR (RT-qPCR) assays, and their secretion products. Using the cell culture medium, maximum cell densities between 0.51 and 0.80 × 10 cells/cm (=2.55-4.00 × 10 cells/mL) were obtained. As the expansion of hASCs represents only the first step in a cell therapeutic protocol or further basic research studies, we formulated two chemically defined media to differentiate the expanded hASCs in white or beige/brown adipocytes. These new media could help translate research projects into the clinical application of hASCs and study ex vivo the biology in healthy and dysfunctional states of adipocytes and their precursors. Following the cell culture system developers' practice and obvious reasons related to the formulas' patentability, the defined media's composition will not be disclosed in this study.

摘要

脂肪组织是干细胞的丰富来源。然而，抽脂术不能获得足够数量的细胞，无法直接应用于再生医学。因此，需要开发符合 GMP 标准的体外扩增方案，以确保生产出安全、可重复且具有成本效益的“细胞药物”。因此，我们开发了自己的基础定义的无动物源和无血清细胞培养液（），专门用于培养人脂肪干细胞（hASCs）。使用这种培养基，我们可以直接在定义的细胞培养条件下培养基质血管部分（SVF）细胞，以获得 hASCs。细胞在保持未分化的情况下增殖，这通过流式细胞术（FACS）、定量逆转录 PCR（RT-qPCR）检测和它们的分泌产物证实。使用 细胞培养液，可以获得 0.51 到 0.80×10 个细胞/cm（=2.55-4.00×10 个细胞/mL）之间的最大细胞密度。由于 hASCs 的扩增仅代表细胞治疗方案或进一步基础研究的第一步，我们还配制了两种化学定义的培养基，以将扩增的 hASCs 分化为白色或米色/棕色脂肪细胞。这些新的培养基可以帮助将研究项目转化为 hASCs 的临床应用，并在体外研究脂肪细胞及其前体细胞在健康和功能障碍状态下的生物学。根据细胞培养系统开发者的实践以及与配方专利相关的明显原因，本研究不会披露定义培养基的组成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f33/7926673/9e4973adb83a/cells-10-00466-g013.jpg
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