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目的建立一种新的造血干细胞获取方法,为临床应用提供实验依据。方法将明胶海绵移植到小鼠后肢的大腿内侧肌间隙,12 d后分离明胶海绵中的迁移细胞。采用流式细胞术和免疫荧光法检测迁移细胞和骨髓细胞中CD34+、Sca-1+和CD34+Sca-1+细胞的表达,利用体外造血克隆分析迁移细胞和骨髓细胞中造血克隆的密度。结果迁移细胞的数量在移植12 d时达最大;迁移细胞中CD34+、Sca-1+和CD34+Sca-1+细胞的比例明显高于骨髓细胞(P<0.05),造血克隆比例也明显高于骨髓(P<0.01)。结论利用生物材料明胶海绵成功建立了一种高效的造血干细胞获取方法,可实现自体细胞治疗。
Objective To establish a new method for obtaining hematopoietic stem cells and provide experimental evidence for clinical application. Methods Gelatin sponge was transplanted into the medial thigh space of the hindlimb of mice. After 12 days, the migrating cells in gelatin sponge were separated. The expression of CD34 +, Sca-1 + and CD34 + Sca-1 + cells in migrating cells and myeloid cells was detected by flow cytometry and immunofluorescence. The hematopoietic clonogenic density in migrating cells and myeloid cells was analyzed by in vitro hematopoietic cloning. Results The number of migrated cells reached the maximum at 12 days after transplantation. The proportion of CD34 +, Sca-1 + and CD34 + Sca-1 + cells in the migrated cells was significantly higher than that in the bone marrow cells (P <0.05), and the proportion of hematopoietic clones was also significantly higher Bone marrow (P <0.01). Conclusion The successful establishment of an efficient method of obtaining hematopoietic stem cells by using biomaterial gelatin sponge can realize autologous cell therapy.