不同来源人造血干/祖细胞在NOD/SCID小鼠体内归巢能力的差异

目的 比较不同来源的人造血干／祖细胞在NOD／SCID小鼠体内归巢能力的差异性，并探讨其体内归巢能力与膜表面归巢相关分子CXCR4表达水平的相关性。方法 采用流式细胞术（FACS）检测新鲜脐血、冻存脐血、动员后外周血（mPB）及骨髓来源的CD34^＋细胞表面CXCR4表达水平；将荧光染料CFSE标记的人CD34^＋细胞移植人接受照射的NOD／SCID小鼠，移植后20小时检测已归巢于NOD／SCID小鼠骨髓及脾脏中不同来源的人CD34^＋细胞，计算其相应的骨髓及脾脏归巢效率；并将小鼠股骨制成组织切片，荧光显微镜下观察人CD34^＋细胞在小鼠骨髓腔内的分布。结果 新鲜脐血、冻存脐血、mPB和骨髓CD34^＋细胞膜表面CXCR4表达阳性率分别为（49.52±1．12）％。（46．12±2．29）％，（48．50±2．48）％和（65．39±1．27）％，CD34^＋细胞在NOD／SCID小鼠骨髓的归巢效率分别为（3．00±0．44）％，（2．84±0．46）％，（4．06±0．70）％及（5．76±0．52）％；在脾脏的归巢率分别为（1．88±0．12）％，（1．80±0．15）％，（1．90±0．22）％，（2．16±0．34）％。归巢的CD34^＋细胞主要分布于小鼠股骨的骨内膜区域。结论 脐血CD34^＋细胞膜表面CXCR4水平低于mPB和骨髓。经冻存复苏后脐血CD34^＋细胞膜表面CXCR4水平略有下调。脐血CD34^＋细胞在照射NOD／SCID小鼠的骨髓归巢效率低于mPB和骨髓。

Differential study on the in vivo homing potential of human hematopoietic stem/progenitor cells from different sources in xenotransplanted NOD/SCID mouse model

OBJECTIVE: To compare the in vivo homing potential of human hematopoietic stem/progenitor cells (HS/PCs) derived from fresh umbilical cord blood (UCB), cryopreserved UCB, mobilized peripheral blood (mPB) and bone marrow (BM) in xenotransplanted NOD/SCID mouse model, and to explore the relationship between the homing potential of HS/PCs and their expression levels of membrane receptor CXCR4. METHODS: The expression levels of membrane CXCR4 on HS/PCs were assessed by flow cytometric analysis (FACS). CFSE-labeled human HS/PCs from different sources were transplanted into irradiated NOD/SCID mice. Human CD34 cells homed in bone marrow and spleen of recipient mice were determined 20 hours after xenotransplantation by FACS and the homing efficiencies were calculated. Tissue sections of the recipient mice femurs were made and the distribution of CFSE-labeled human CD34 cells were observed under fluorescence microscope. RESULTS: The expression levels of membrane CXCR4 on CD34+ cells from fresh UCB, cryopreserved UCB, mPB and BM were (49.52 +/- 1.12)%, (46.12 +/- 2.29)%, (48.50 +/- 2.48)% and (65.39 +/- 1.27)%, respectively. The homing efficiencies of CD34+ cells from fresh UCB, cryopreserved UCB, mPB and BM in recipient mice BM were (3.00 +/- 0.44)%, (2.84 +/- 0.46)%, (4.06 +/- 0.70)% and (5.76 +/- 0.52)% , respectively. Human CD34+ cells mainly located within endosteal region of recipient mice femurs. CONCLUSION: CD34+ cells from UCB express lower levels of membrane CXCR4 than those from mPB and BM. The level of membrane CXCR4 on UCB CD34+ cells is down-regulated after freezing and thawing procedures. The homing efficiency of human CD34 cells from UCB in recipient mice is lower than that of mPB and BM.