骨髓腔输注异基因间充质干细胞对移植后大鼠骨髓间充质干细胞重建的作用

为了研究骨髓腔内注射（IBM）异基因间充质干细胞（MSC）对造血干细胞移植后大鼠骨髓MSC重建的作用，研究供体MSC植入状态以探讨MSCs的作用机制，将雌性F344胎鼠及新生鼠外周血（FNPB）及雄性F344大鼠BrdU标记骨髓MSC共移植入经致死量^60Coγ射线预处理的雌性Wistar大鼠，其中FNPB均由IBM输注，MSC则通过IBM或尾静脉注射。观察受鼠存活状况、供体HSC植入水平及骨髓MSC恢复情况，并以PCR检测Y染色体和免疫荧光法检测受鼠骨髓MSC的来源。结果显示：两个FNPB＋MSCs共移植组大鼠移植后60天均100％存活，两组比较生存率和供体HSC植入水平无统计学差异，但明显优于单纯FNPB移植组；移植后30天时各移植组受鼠骨髓MSC的增殖能力均未达正常水平，但MSC骨髓腔共移植组集落数（66．0±10．6）明显优于MSC骨髓腔和静脉共移植组及单纯FNPB移植组（P〈0．01）；移植后60天时，免疫荧光法检测供体BrdU标记的MSC显示仅在少部分受体发现供、受体源性MSCs嵌合，但两个共移植组存活受鼠均检测到供体MSC来源Y染色体。结论：异基因MSC输注可促进造血干细胞移植受者骨髓MSC恢复，尤以IBM输注为佳。

Effect of Intra-Bone Marrow Infusion of Allogeneic Mesenchymal Stem Cells on Reconstruction of Marrow Mesenchymal Stem Cells in Rat HSCT nodels

This study was aimed to investigate the effect of intra-bone marrow （IBM） injection of aUogeneic mesenchymal stem cells （MSCs） on reconstruction of bone marrow MSCs （BM-MSCs） in rats that received hematopoietic stem cell transplantation （ HSCT）, and to detect the donor MSCs in the hosts for clarifying the effect mechanism of donor MSCs. Wistar female rats conditioned with lethal dose ^60Co γ-ray irradiation were co-transplanted with F344 female fetal and neonatal peripheral blood （FNPB） and BrdU-labeled MSCs separated from bone marrow mononuclear cells of F344 male rats. The donor MSCs were infused by IBM injection in bilateral tibia or intravenous injection （ Ⅳ）, while the FNPB were all via IBM route. The survival rate, engraftment level of HSCs and recovery of BM-MSCs of recipients were monitored. The ratio of BrdU-labeled MSCs in recipient rats was calculated by immunofluorescence assay （IFA） and the Y chromosomes were examined by PCR. The results showed that the recipient rats of the two co-transplantation groups were all alive at day 60 after tranplantation. There was no significant difference between these two groups on the survival rates or the engraftment levels of HSCs, but each of them was much better than that of the FNPB group. At day 30 after transplantation, the proliferation ability of recipients＇BM-MSCs was still below normal, while that of the FNPB （IBM） ＋ MSC（IBM） group was the best of all the experiment groups（p 〈 0.01 ）. At 60 days, the donor MSCs coexisted with host MSCs in only a few recipient rats examined by IFA, while the Y chromosomes could be detected in all the recipient rats in the two cotransplantation groups. It is concluded that the infusion of allogeneic MSCs can accelerate the recovery of HSCT recipients＇BM-MSCs. The IBM route is safe and more effective than infravenous infusion.