Freitas Gileade P, Souza Alann T P, Lopes Helena B, Trevisan Rayana L B, Oliveira Fabiola S, Fernandes Roger R, Ferreira Fernanda U, Ros Felipe A, Beloti Marcio M, Rosa Adalberto L
Bone Research Lab, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
Hemotherapy Center of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
Bio Protoc. 2020 Feb 20;10(4):e3534. doi: 10.21769/BioProtoc.3534.
Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and regenerative medicine. In this article, we describe protocols to isolate MSCs from bone marrow (BM-MSCs) and adipose tissues (AT-MSCs), and methods to culture, characterize, and differentiate MSCs into osteoblasts, adipocytes, and chondrocytes. After the harvesting of cells from bone marrow by flushing the femoral diaphysis and enzymatic digestion of abdominal and inguinal adipose tissues, MSCs are selected by their adherence to the plastic tissue culture dish. Within 7 days, MSCs reach 70% confluence and are ready to be used in subsequent experiments. The protocols described here are easy to perform, cost-efficient, require minimal time, and yield a cell population rich in MSCs.
自间充质基质细胞（MSCs）被发现以来，它们受到了广泛关注，主要是由于其自我更新潜力和多向分化能力。基于这些原因，MSCs在细胞生物学和再生医学中是一种有用的工具。在本文中，我们描述了从骨髓（BM-MSCs）和脂肪组织（AT-MSCs）中分离MSCs的方案，以及培养、鉴定MSCs并将其分化为成骨细胞、脂肪细胞和软骨细胞的方法。通过冲洗股骨干采集骨髓细胞以及对腹部和腹股沟脂肪组织进行酶消化后，通过MSCs对塑料组织培养皿的贴壁特性进行筛选。7天内，MSCs达到70%汇合度，即可用于后续实验。这里描述的方案易于操作、成本效益高、所需时间最少，并且能产生富含MSCs的细胞群体。
