冷冻保存对人脂肪组织和分离的基质血管部分细胞的影响：体外和体内分析。
Effect of Cryopreservation on Human Adipose Tissue and Isolated Stromal Vascular Fraction Cells: In Vitro and In Vivo Analyses.
机构信息
São Paulo and Brasilia, Brazil; and New Orleans, Covington, and Baton Rouge, La.
From the Translational Surgery Graduate Program and Plastic Surgery Division, Federal University of São Paulo; the Center for Stem Cell Research & Regenerative Medicine, Tulane University; the Department of Cell and Molecular Biology, Tulane University School of Science and Engineering; the Physics Department, Dillard University; La Cell LLC; the Departments of Pharmacology, Medicine, Structural and Cellular Biology, and Surgery, Tulane University School of Medicine; the Tulane National Primate Research Center; Plastic Surgery Associates; the Department of Mechanical Engineering, Louisiana State University; and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.
出版信息
Plast Reconstr Surg. 2018 Feb;141(2):232e-243e. doi: 10.1097/PRS.0000000000004030.
BACKGROUND
Adipose tissue is a source of adipose-derived stromal/stem cells for tissue engineering and reconstruction and a tissue source for fat grafts. Although liposuction is a simple procedure for the harvest of adipose tissue, the repetition of this surgical intervention can cause adverse effects to the patient and can be a limiting factor for immediate use. Cryopreservation can avoid the morbidity associated with repetitive liposuction, allowing the use of stored tissue after the initial harvest procedure. This article focuses on the characterization of fresh and cryopreserved human adipose tissue.
METHODS
Lipoaspirates from eight donors were processed as fresh adipose tissue or cryopreserved for 4 to 6 weeks. Fresh and cryopreserved tissues were collagenase digested and the stromal vascular fraction cells were characterized immediately or cryopreserved. Characterization was based on stromal vascular fraction cell proliferation and immunophenotype. In vivo fat grafting was performed in C57BL/6 green fluorescent protein mice to analyze morphology of the tissue and its adiposity using confocal microscopy, histochemical staining (i.e., hematoxylin and eosin and Masson trichrome), and immunohistochemistry (i.e., green fluorescent protein, perilipin, and CD31).
RESULTS
Although tissue and stromal vascular fraction cell cryopreservation reduced the total cell yield, the remaining viable cells retained their adhesive and proliferative properties. The stromal vascular fraction cell immunophenotype showed a significant reduction in the hematopoietic surface markers and increased expression of stromal and adipogenic markers following cryopreservation. In vivo cryopreserved fat grafts showed morphology similar to that of freshly implanted fat grafts.
CONCLUSION
In this study, the authors demonstrated that cryopreserved adipose tissue is a potential source of stromal vascular fraction cells and a suitable source for fat grafts.
背景
脂肪组织是组织工程和重建的脂肪来源基质/干细胞的来源，也是脂肪移植物的组织来源。虽然吸脂术是一种简单的收获脂肪组织的方法，但这种手术干预的重复会对患者造成不良影响，并且可能成为即时使用的限制因素。冷冻保存可以避免与重复吸脂相关的发病率，允许在初始收获程序后使用储存的组织。本文重点介绍新鲜和冷冻保存的人脂肪组织的特性。
方法
从 8 名供体中提取的脂肪抽吸物被加工为新鲜脂肪组织或冷冻保存 4 至 6 周。新鲜和冷冻保存的组织用胶原酶消化，基质血管成分细胞立即或冷冻保存后进行特征描述。特征描述基于基质血管成分细胞的增殖和免疫表型。将 C57BL/6 绿色荧光蛋白小鼠进行体内脂肪移植，使用共聚焦显微镜、组织化学染色（即苏木精和伊红、马松三色染色）和免疫组织化学（即绿色荧光蛋白、脂联素和 CD31）分析组织的形态及其脂肪含量。
结果
尽管组织和基质血管成分细胞的冷冻保存降低了总细胞产量，但剩余的活细胞仍保持其黏附和增殖特性。基质血管成分细胞的免疫表型显示，冷冻保存后造血表面标志物显著减少，而基质和脂肪生成标志物的表达增加。体内冷冻保存的脂肪移植物的形态与新鲜植入的脂肪移植物相似。
结论
在这项研究中，作者证明了冷冻保存的脂肪组织是基质血管成分细胞的潜在来源，也是脂肪移植物的合适来源。
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