Department of Biochemistry and Structural Biology, University of Texas Health Science Center.
Department of Biomedical Engineering and Chemical Engineering, University of Texas at San Antonio; Institute of Regenerative Medicine, University of Texas at San Antonio.
J Vis Exp. 2023 Feb 3(192). doi: 10.3791/64650.
Engineering thermogenic adipose tissue (e.g., beige or brown adipose tissues) has been investigated as a potential therapy for metabolic diseases or for the design of personalized microtissues for health screening and drug testing. Current strategies are often quite complex and fail to accurately fully depict the multicellular and functional properties of thermogenic adipose tissue. Microvascular fragments, small intact microvessels comprised of arteriole, venules, and capillaries isolated from adipose tissue, serve as a single autologous source of cells that enable vascularization and adipose tissue formation. This article describes methods for optimizing culture conditions to enable the generation of three-dimensional, vascularized, and functional thermogenic adipose tissues from microvascular fragments, including protocols for isolating microvascular fragments from adipose tissue and culture conditions. Additionally, best practices are discussed, as are techniques for characterizing the engineered tissues, and sample results from both rodent and human microvascular fragments are provided. This approach has the potential to be utilized for the understanding and development of treatments for obesity and metabolic disease.
工程化生热脂肪组织（如米色或棕色脂肪组织）已被研究作为代谢疾病的潜在治疗方法，或用于设计个性化微组织以进行健康筛查和药物测试。目前的策略通常非常复杂，并且无法准确地充分描绘出生热脂肪组织的多细胞和功能特性。微血管片段是从小鼠脂肪组织中分离出来的由小动脉、小静脉和毛细血管组成的完整的微小血管，作为一种能够实现血管生成和脂肪组织形成的自体细胞的单一来源。本文描述了优化培养条件的方法，以从微血管片段中生成三维、血管化和功能性的生热脂肪组织，包括从脂肪组织中分离微血管片段的方案和培养条件。此外，还讨论了最佳实践，以及用于表征工程化组织的技术，并且提供了来自啮齿动物和人类微血管片段的样本结果。这种方法有可能被用于理解和开发肥胖和代谢疾病的治疗方法。
