Human adipose tissue includes useful substrates for regenerative medicine such as the extracellular matrix (ECM), but most perirenal fat tissue is wasted after kidney surgery. Since a lot of adipose tissue can be procured after a kidney, we extracted ECM from human perirenal adipose tissue and optimized the extraction process. To verify the efficacy for ECM extraction, we compared the products in several steps. Perirenal adipose tissue was either finely homogenized or underwent crude manual dissection. The amount of extracted ECM was quantified with ELISA for verification of the initial tissue downsizing effect. To validate the drying effect for fast and complete delipidation, tissues were prepared in a dry or wet phase, and residual lipids were visualized with Oil-Red-O staining. The extracted lipid was assayed at each time point to quantify the appropriate delipidation time. To select the optimal decellularization method, tissues were treated with physical, chemical, or enzymatic method, and the residual cell debris were identified with histological staining. The biochemical properties of the ECM extracted by the above methods were analyzed. The ECM extracted by fine homogenization showed a significantly enhanced amount of collagen, laminin and fibronectin compared to the crude dissection method. The dried tissue showed fast and complete lipid elimination compared to the wet tissue. Complete delipidation was achieved at 45 min after acetone treatment. Additionally, 1% triton X-100 chemical treatment showed complete decellularization with well-preserved collagen fibers. Biochemical analysis revealed preserved ECM proteins, a high cell proliferation rate and normal cell morphology without cell debris or lipids. The established process of homogenization, drying, delipidation with acetone, and decellularization with Triton X-100 treatment can be an optimal method for ECM extraction from human perirenal adipose tissue. Using this technique, human perirenal adipose tissue may be a valuable source for tissue engineering and regenerative medicine.

人体脂肪组织包括用于再生医学的有用基质，例如细胞外基质 (ECM)，但大多数肾周脂肪组织在肾脏手术后被浪费。由于肾脏后可以获取大量脂肪组织，我们从人肾周脂肪组织中提取了 ECM，并优化了提取过程。为了验证 ECM 提取的功效，我们分几个步骤比较了产品。肾周脂肪组织要么精细均质化，要么进行粗略的手动解剖。用 ELISA 对提取的 ECM 的量进行量化，以验证初始组织缩小效果。为了验证快速和完全脱脂的干燥效果，在干或湿相中制备组织，并用油红 O 染色观察残留的脂质。在每个时间点测定提取的脂质以量化适当的脱脂时间。为了选择最佳的脱细胞方法，组织用物理、化学或酶法处理，残留的细胞碎片用组织学染色鉴定。对上述方法提取的ECM的生化性质进行了分析。与粗解剖方法相比，通过精细均质提取的 ECM 显示出胶原蛋白、层粘连蛋白和纤连蛋白的含量显着增加。与湿组织相比，干组织显示出快速和完全的脂质消除。在丙酮处理后 45 分钟实现完全脱脂。此外，1% triton X-100 化学处理显示完全脱细胞，胶原纤维保存完好。生化分析显示保存的 ECM 蛋白，细胞增殖率高，细胞形态正常，无细胞碎片或脂质。已建立的均质化、干燥、用丙酮脱脂和用 Triton X-100 处理去细胞化的过程可能是从人肾周脂肪组织中提取 ECM 的最佳方法。使用这种技术，人类肾周脂肪组织可能是组织工程和再生医学的宝贵来源。