Objective: To establish an intramedullary transplantation model of primary megakaryocytes to evaluate the platelet-producing capacity of megakaryocytes and explore the underlying regulatory mechanisms. Methods: Donor megakaryocytes from GFP-transgenic mice bone marrow were enriched by magnetic beads. The platelet-producing model was established by intramedullary injection to recipient mice that underwent half-lethal dose irradiation 1 week in advance. Donor-derived megakaryocytes and platelets were detected by immunofluorescence staining and flow cytometry. Results: The proportion of megakaryocytes in the enriched sample for transplantation was 40 to 50 times higher than that in conventional bone marrow. After intramedullary transplantation, donor-derived megakaryocytes successfully implanted in the medullary cavity of the recipient and produce platelets, which showed similar expression of surface markers and morphology to recipient-derived platelets. Conclusion: We successfully established an in vivo platelet-producing model of primary megakaryocytes using magnetic-bead enrichment and intramedullary injection, which objectively reflects the platelet-producing capacity of megakaryocytes in the bone marrow.
目的： 利用小鼠原代巨核细胞骨髓腔移植构建血小板产生模型，为研究巨核细胞功能及血小板产生调控机制提供工具。 方法： 通过磁珠富集绿色荧光蛋白（Green Fluorescent Proteins, GFP）转基因供体小鼠骨髓原代巨核细胞，将其移植入经半致死剂量辐照的受体小鼠骨髓腔，建立原代巨核细胞骨髓腔内血小板生成模型。通过免疫荧光染色、流式细胞术等方法检测受体小鼠中供体来源巨核细胞和血小板形态、大小等指标。 结果： 磁珠富集可将巨核细胞在骨髓细胞悬液中的比例提高40~50倍。供体来源巨核细胞能够在受体小鼠骨髓腔成功定植并正常产生血小板，其产生的子代血小板具有与受体自身来源血小板相似的形态、大小及CD41、CD42d、CD61等表面标志分子表达水平。 结论： 通过磁珠富集与骨髓腔注射可成功构建小鼠原代巨核细胞移植模型，该模型能够客观反映巨核细胞在骨髓内产生血小板的能力。.
Keywords: Intramedullary injection; Megakaryocyte; Platelet.