State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University.
College of Life Sciences, Northwest University.
J Vis Exp. 2022 Apr 13(182). doi: 10.3791/63811.
Human mesenchymal stem cells (hMSCs) have shown great potential in bone regeneration, immune modulation, and treating refractory chronic diseases. Various origins have been found to obtain hMSCs recently, while bone marrow was still considered the main source. Bone marrow-derived MSCs (BMSCs) from different donor bone sites have distinct characteristics due to microenvironmental factors. Studies have shown that BMSCs from maxillofacial bone may have greater proliferative and osteogenic capacities than BMSCs from long bones or the iliac crest. And maxillofacial BMSCs were considered more suitable for stem cell therapy in the maxillofacial tissues. The mandible, especially the ascending ramal area with sufficient marrow, was a feasible donor site for harvesting BMSCs. This study described a protocol for harvesting, isolating, and culturing human mandibular bone marrow-derived MSCs (hmBMSCs). Furthermore, immunophenotyping of hmBMSCs, proliferation assays, and in vitro induction of osteogenic, adipogenic, and chondrogenic differentiation was performed to identify the cultured cells. Applying this protocol can help the researchers successfully obtain enough high-quality hmBMSCs, which is necessary for further studies of the biological function, microenvironmental effects, and clinical applications.
人骨髓间充质干细胞（hMSCs）在骨再生、免疫调节和治疗难治性慢性疾病方面显示出巨大的潜力。最近发现了各种来源来获取 hMSCs，而骨髓仍然被认为是主要来源。由于微环境因素的影响，来自不同供体骨部位的骨髓间充质干细胞（BMSCs）具有不同的特征。研究表明，来自颌面骨的 BMSCs 比来自长骨或髂嵴的 BMSCs 具有更大的增殖和成骨能力。而且，颌面 BMSCs 被认为更适合用于颌面组织的干细胞治疗。下颌骨，特别是有足够骨髓的升支区域，是采集 BMSCs 的可行供体部位。本研究描述了一种从人下颌骨骨髓中采集、分离和培养间充质干细胞（hmBMSCs）的方案。此外，对 hmBMSCs 进行免疫表型分析、增殖试验以及体外诱导成骨、成脂和成软骨分化，以鉴定培养的细胞。应用该方案有助于研究人员成功获得足够数量的高质量 hmBMSCs，这对于进一步研究其生物学功能、微环境影响和临床应用是必要的。
