脊柱手术出血中间充质干细胞的分离鉴定及数量变化*★

背景：胰肾联合移植已经被公认为是糖尿病(包括1型和2型)合并终末期尿毒症的有效治疗手段，由于胰腺为高免疫原性器官，合理的免疫抑制治疗是保证胰腺移植成功的关键。 目的： 探讨胰肾一期联合移植后免疫抑制药物的合理应用。 方法：纳入2005-01/2009-06在中山大学附属第一医院器官移植中心完成胰肾一期联合移植的患者9例，其中男5例，女4例，胰液引流均采用空肠引流方式。术后采用白细胞介素2单克隆抗体诱导的四联免疫抑制方案：白细胞介素2单克隆抗体+他克莫司+麦考酚酸+激素，并逐渐过渡至单用他克莫司维持治疗。回顾性分析以上9例患者围手术期及长期随访情况。 结果与结论：胰肾一期联合移植后，除1例早期死亡外，其余8例患者移植后1周内肌酐降至正常水平，移植后停用胰岛素时间为(11.5±3.5) d，空腹血糖恢复至正常时间为(15.4±6.3) d。8例患者随访4~50个月期间，共有4例发生移植肾急性排斥，其中1例在接受床边血液透析过程中并发心脑血管意外后家属放弃治疗，其余3例患者经抗胸腺细胞球蛋白或激素冲击治疗后移植肾功能均逆转恢复，随访过程中未发现移植胰腺排斥。说明胰肾联合移植是治疗糖尿病合并终末期糖尿病肾病的有效方法，术后早期采用白细胞介素2单克隆抗体诱导的四联免疫抑制方案并逐渐过渡至单用他克莫司维持治疗是安全的。

A preliminary study of the number and identification of mesenchymal stem cells in spinal surgery bleeding

BACKGROUND: A great amount of red bone marrow in the cancellous bone losses during orthopedic surgery. OBJECTIVE: To explore the feasibility to isolate bone marrow mesenchymal stem cells (BMSCs) during spinal surgery bleeding and to investigate the relationship of the number of BMMSCs with age and time. METHODS: Three cases of degenerative lumbar instability and 16 cases of lumbar burst fractures were selected to undergo posterior pedicle screw fixation, including 4 cases of 20-30 years, 5 of 30-40 years, 5 of 40-50 years, 2 of 50-60 years, 1 of 64 years, 1 of 65 years, and 1 of 67 years. 5 mL of vertebra blood was collected from 4 cases within 0-5 minutes after the awl for pierced vertebra was pulled out, and 5 mL of vertebral blood from other 15 cases was collected within 0-5, 5-10 and 10-15 minutes after the awl was pulled out, respectively. Mononuclear cells were isolated and incubated into 6-well plates, and then cultured for 3 days. The medium was renewed completely to remove unadherent cells. Hematoxylin-eosin staining was applied for cell counting. RESULTS AND CONCLUSION: The membrane mark showed that CD44 and CD29 were positive while the CD34 and CD14 were negative. The number of BMSCs isolated within 0-5 minutes was 262.47±172.20, and fewer or no BMSCs were found within 5-10 or 10-15 minutes. The number of BMMSCs from vertebra bleeding of 0-5 minutes had a decreased tendency with aging.