去细胞脂肪组织：通过招募和诱导间充质干细胞促进脂肪再生的关键因素。
Decellularized adipose tissue: A key factor in promoting fat regeneration by recruiting and inducing mesenchymal stem cells.
机构信息
Department of Plastic Surgery, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong Province, 510630, PR China; Innovative Technology Research Institute of Tissue Repair and Regeneration, Key Laboratory of Regenerative Medicine, Ministry of Education, Guangzhou, Guangdong Province, 510630, PR China.
出版信息
Biochem Biophys Res Commun. 2021 Feb 19;541:63-69. doi: 10.1016/j.bbrc.2020.12.108. Epub 2021 Jan 19.
BACKGROUND
Decellularized adipose tissue (DAT) has attracted much attention due to its wide range of sources and adipose regeneration capacity. However, the lipogenic efficiency of DAT is still controversial due to its unclear mechanism. To this point, it is crucial to clarify the mechanism of DAT in promoting adipose regeneration Objective: This study aims to explore the mechanism of DAT promoting adipose regeneration and survival mechanism of DAT transplantation in vivo.
METHODS
DAT preparation by repeated freeze-thaw, enzymatic digestion, and isopropanol degreasing. Histology, DAPI, immunohistochemistry, immunofluorescence and scanning electron microscopy confirmed the efficacy and reproducibility of these approaches. BM-MSCs, ADSCs and UCMSCs were cocultured with DAT for 14 days and then stained with oil red O. Adipogenic genes of three MSCs were detected by RT-PCR. DAT and adipose tissue were transplanted subcutaneously into the back of nude mice to observe medium and long-term morphological changes, vascularization, and lipid-forming efficiency. Mass spectrometry (MS)-based proteomic to analyze the adipogenic protein contents of DAT and adipose tissue.
RESULTS
The DAT without any cellular components but with an abundance of collagen; neither DNA nor lipids were detected. Seeding experiments with MSCs indicated that the DAT provided an inductive microenvironment for adipogenesis, supporting the expression of the master regulators PPARγ. Within four months after transplantation, HE morphology of DAT was identical to adipose cells. Immunofluorescence markers CD31 and perilipin were increased in DAT, while the retention rate gradually decreased over time, eventually accounting for 33.7% of the original volume. MS-based proteomic analyses identified 1013 types of proteins in adipose tissue and 29 proteins in the DAT. Analyses of GO and KEGG databases suggested that DAT contained a variety of proteins involved in fat metabolism.
CONCLUSIONS
DAT can interact with different types of MSCs and ultimately achieve adipose regeneration. The presence of multiple adipogenic proteins in DAT make it play a vital role in adipose regeneration. DAT is expected to be an ideal bio-derived scaffold for adipose tissue engineering.
背景
去细胞脂肪组织（DAT）因其来源广泛和脂肪再生能力而受到广泛关注。然而，由于其机制不清楚，DAT 的脂肪生成效率仍存在争议。在这一点上，阐明 DAT 促进脂肪再生的机制和 DAT 体内移植的存活机制至关重要。
目的
本研究旨在探讨 DAT 促进脂肪再生的机制和 DAT 体内移植的存活机制。
方法
通过反复冻融、酶消化和异丙醇脱脂制备 DAT。组织学、DAPI、免疫组织化学、免疫荧光和扫描电子显微镜证实了这些方法的有效性和可重复性。将 BM-MSCs、ADSCs 和 UCMSCs 与 DAT 共培养 14 天，然后用油红 O 染色。通过 RT-PCR 检测三种 MSC 的脂肪生成基因。将 DAT 和脂肪组织皮下移植到裸鼠背部，观察中、长期形态变化、血管生成和脂质形成效率。基于质谱（MS）的蛋白质组学分析 DAT 和脂肪组织中的脂肪生成蛋白含量。
结果
DAT 不含任何细胞成分，但含有丰富的胶原蛋白；既没有检测到 DNA 也没有检测到脂质。MSC 接种实验表明，DAT 为脂肪生成提供了诱导微环境，支持主调控因子 PPARγ 的表达。移植后 4 个月，DAT 的 HE 形态与脂肪细胞相同。免疫荧光标志物 CD31 和 perilipin 在 DAT 中增加，而保留率随时间逐渐降低，最终占原始体积的 33.7%。基于 MS 的蛋白质组学分析鉴定出脂肪组织中有 1013 种蛋白质和 DAT 中有 29 种蛋白质。GO 和 KEGG 数据库的分析表明，DAT 含有多种参与脂肪代谢的蛋白质。
结论
DAT 可以与不同类型的 MSC 相互作用，最终实现脂肪再生。DAT 中存在多种脂肪生成蛋白使其在脂肪再生中发挥重要作用。DAT 有望成为脂肪组织工程的理想生物衍生支架。
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