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目的探讨人脐血源基质细胞(human umbilical cord blood-derived stromal cells,hUCBDSCs)对造血细胞早期归巢及植入能力的影响。方法 CCK-8法和Transwell法分别比较hUCBDSCs和人骨髓基质细胞(human bone marrow stromal cells,hBMSCs)对脐血单个核细胞(human umbilical cord blood-mononuclear cells,hUCB-MNCs)体外黏附及迁移能力的影响;RT-PCR法检测hUCBDSCs对促归巢相关因子的表达;60Coγ射线致死剂量辐照BABL/c小鼠后分别接受hUCB-MNCs单移植或hUCB-MNCs联合基质细胞共移植,流式细胞仪检测小鼠骨髓人源CD45+细胞植入率;追踪移植后CM-DiI标记的hUCB-MNCs体内迁移情况,并比较各组归巢率。结果 hUCBDSCs和hBMSCs均能促进hUCB-MNCs的黏附,且迁移作用显著强于无基质细胞共培养对照(P<0.05);hUCBDSCs能表达一系列与造血归巢相关的黏附分子和细胞因子;hUCBDSCs在联合hUCB-MNCs共移植中能不同程度提高造血植入率,且当输注的hUCB-MNCs为低剂量(两种细胞按1∶1各移植2×106/只)时这种促进作用最为显著(P<0.01);CM-DiI标记的hUCB-MNCs在体内主要归巢至骨髓和脾脏,hUCBDSCs联合移植后48 h归巢率(43.49±3.06)%显著高于hBMSCs联合移植组[(31.99±7.26)%]和单移植组[(21.46±6.88)%],(P<0.05)。结论 hUCBDSCs在促进造血细胞迁移、归巢和植入方面作用显著。
Objective To investigate the effect of human umbilical cord blood-derived stromal cells (hUCBDSCs) on early homing and implantation of hematopoietic cells. Methods The adhesion and migration of hUCBDSCs and human bone marrow stromal cells (hBMSCs) to human umbilical cord blood-mononuclear cells (hUCB-MNCs) in vitro were compared between CCK-8 and Transwell methods HUCBDSCs were detected by RT-PCR and BABL / c mice were irradiated with 60Coγ-ray lethal dose respectively. The mice were either transplanted with hUCB-MNCs alone or hUCB-MNCs combined with stromal cells. Flow cytometry The implantation rate of murine bone marrow-derived CD45 + cells was detected. The migration of CM-Di-labeled hUCB-MNCs in vivo was followed up and the homing rate of each group was compared. Results Both hUCBDSCs and hBMSCs could promote the adhesion of hUCB-MNCs, and the migration of hUCBDSCs was significantly stronger than that of the control (P <0.05). HUCBDSCs could express a series of adhesion molecules and cytokines related to hematopoietic homing. In the co-transplantation of hUCB-MNCs, the rate of hematopoietic engraftment could be improved to some extent, and this effect was the most significant when the infused hUCB-MNCs were low dose (2 × 10 6 cells per 1: 1 for both cells) (P <0.01). The hUCB-MNCs labeled with CM-DiI mainly homing to the bone marrow and spleen in vivo. The homing rate (43.49 ± 3.06)% at 48 h after hUCBDSCs transplantation was significantly higher than that in hBMSCs transplantation group [(31.99 ± 7.26)%] and single transplantation group [(21.46 ± 6.88)%], (P <0.05). Conclusion hUCBDSCs play a significant role in promoting hematopoietic cell migration, homing and implantation.