摘要目的 探讨联合成骨细胞移植对骨髓移植小鼠造血功能重建的影响.方法 Balb/c小鼠60只,取其中18只制备移植用骨髓有核细胞和成骨细胞.将其余42只小鼠分为3组.单纯移植组:小鼠18只,仅进行骨髓移植(BMT);联合移植组:18只,进行BMT的同时每只小鼠输入成骨细胞2×106个;正常对照组:小鼠6只,不做任何处理,仅作为移植前的正常对照.移植组小鼠在全身照射(TBI)预处理后4 h经尾静脉注入骨髓细胞2×106个.移植后第7、14和21天,分别处死移植组小鼠6只,对小鼠的外周血细胞和骨髓单个核细胞(BMMNC)进行计数,采用流式细胞术测定BMMNC中CD34+细胞的百分比,使用HPIAS-1000高清晰度彩色病理图像系统测量骨髓造血组织面积,采用免疫组化染色法测定骨髓组织微血管密度(MVD).结果 移植后第7天,单纯移植组和联合移植组小鼠外周血细胞计数及BMMNC数均明显低于正常对照(P＜0.01).第21天时联合移植组小鼠白细胞、红细胞及BMMNC数明显恢复,血小板数已接近正常对照组;单纯移植组小鼠外周血细胞数和BMMNC数虽然有所恢复,但其恢复程度明显弱于联合移植组.移植后第7、14和21天,联合移植组外周血细胞计数及BMMNC数均明显高于同期单纯移植组(P＜0.01或P＜0.05).移植后第7、14、21天,单纯移植组和联合移植组小鼠骨髓造血组织面积、BMMNC中CD34+细胞百分比及骨髓组织MVD均明显低于正常对照组(P＜0.01),但联合移植组均高于同期单纯移植组(P＜0.01或P＜0.05).结论 联合成骨细胞移植能有效促进骨髓移植小鼠骨髓造血系统的重建.

abstractsObjective To explore the effects of cotransplantation with osteoblasts on hematopoietic reconstitution in mice after bone marrow transplantation (BMT). Methods The typical model of syngeneic BMT was established. 18 Balb/c mice were used to prepare the bone marrow nuclear cells and osteoblasts for BMT. The 42 Balb/c mice were randomly divided into 3 group:normal group (6 mice, without any treatment), the single BMT group ( 18 mice, given 2 × 106 bone marrow nuclear cells/each mouse) and the cotransplantation group of HSC with osteoblaats (18 mice,given 2 × 106 bone marrow nuclear cells and osteoblasts/each mouse). The following factors were measured on day 7, 14, 21 after BMT: peripheral blood cells, bone marrow mononuclear cells (BMMNC), the percentage of CD34+ cells in BMMNC (assayed by flow cytometry), the hematopoietic tissue changes (detected by HPIAS-1000 image analysis system) and micro vascular density (MVD) of bone marrow tissue (with immunohistochemistry). Results The levels of periphral WBC, RBC, PLT, BMMNC in the contransplantation group were higher than those in the single BMT group (P＜0. 01 or P＜0. 05). In the contransplantation group, the percentage of CD34+ cells in BMMNC, the hematopoietic tissue area and the MVD of bone marrow were also higher than the single BMT group on the 7th, 14th, 21st day after BMT(P＜0.01 or P＜0.05). Conclusion Cotransplantation with osteoblasts could significantly promote hematopoietic reconstruction in mice after BMT. Cotransplantation may represent a promising means of achieving higher engraftment rate after BMT.