Yang Zhou, Lu Shi-hong, Li Yan-han, Liu Bin, Wang Hui-jun, Jia Hai-rong, Zheng Yi-zhou
State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC, Tianjin 300020, China.
Zhonghua Xue Ye Xue Za Zhi. 2007 Jun;28(6):391-5.
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.
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 home 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.
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.
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.
在异种移植的NOD/SCID小鼠模型中比较源自新鲜脐带血(UCB)、冷冻保存的UCB、动员外周血(mPB)和骨髓(BM)的人造血干/祖细胞(HS/PCs)的体内归巢潜能,并探讨HS/PCs归巢潜能与其膜受体CXCR4表达水平之间的关系。
通过流式细胞术分析(FACS)评估HS/PCs上膜CXCR4的表达水平。将来自不同来源的CFSE标记的人HS/PCs移植到经照射的NOD/SCID小鼠体内。异种移植20小时后,通过FACS测定受体小鼠骨髓和脾脏中归巢的人CD34细胞,并计算归巢效率。制作受体小鼠股骨的组织切片,在荧光显微镜下观察CFSE标记的人CD34细胞的分布。
新鲜UCB、冷冻保存的UCB、mPB和BM中CD34+细胞上膜CXCR4的表达水平分别为(49.52±1.12)%、(46.12±2.29)%、(48.50±2.48)%和(65.39±1.27)%。新鲜UCB、冷冻保存的UCB、mPB和BM中CD34+细胞在受体小鼠骨髓中的归巢效率分别为(3.00±0.44)%、(2.84±0.46)%、(4.06±0.70)%和(5.76±0.52)%。人CD34+细胞主要位于受体小鼠股骨的骨内膜区域。
UCB来源的CD34+细胞比mPB和BM来源的细胞表达更低水平的膜CXCR4。冷冻和解冻过程后,UCB CD34+细胞上的膜CXCR4水平下调。UCB来源的人CD34细胞在受体小鼠中的归巢效率低于mPB和BM来源的细胞。