Yang Shi, Cai Hai-Bo, Jin Hui-Li, Tan Wen-Song
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Oct;24(10):947-9.
To study the hematopoietic reconstitution ability of fresh and cultured cord blood CD34(+) cells by sublethally irradiated NOD/SCID mice.
Mononuclear cells (MNC) were separated from cord blood and then cultured with stem cell factor (SCF)+ flt3 ligand (FL)+ thrombopoietin (TPO)+ interleukin-3 (IL-3)+ IL-6 for 14 d. CD34(+) cells were isolated from fresh or cultured MNC using miniMACS magnetic separation system. Then 4x10(5) CD34(+) cells together with 5x10(6) CD34(-) cells were injected into NOD/SCID mice. After transplantation, the dynamics of hematopoieitc recovery in the mice were measured. After 6 weeks, bone marrow (BM) and spleen samples were obtained from mice for detecting human cells.
The number of total cells increased 1.78-fold after 14 d of culture. All recipients received fresh and cultured CD34(+) cells survived. Human cells, human lineage cells and human ALU sequence could be detected in BM and spleen. The peripheral blood counts of all transplanted mice recovered. The engraftment levels of cultured CD34(+) cells were similar to those of fresh CD34(+)cells but the percentage of human lineage cells was higher than that of fresh CD34(+) cells.
CD34(+) cells cultured for 14 d still preserved the hematopoietic reconstitution ability and showed better multilineage reconstitution ability than fresh CD34(+) cells.
通过亚致死剂量照射的NOD/SCID小鼠研究新鲜和培养的脐血CD34(+)细胞的造血重建能力。
从脐血中分离单个核细胞（MNC），然后用干细胞因子（SCF）+fms样酪氨酸激酶3配体（FL）+血小板生成素（TPO）+白细胞介素-3（IL-3）+IL-6培养14天。使用miniMACS磁性分离系统从新鲜或培养的MNC中分离CD34(+)细胞。然后将4×10(5)个CD34(+)细胞与5×10(6)个CD34(-)细胞一起注入NOD/SCID小鼠。移植后，检测小鼠造血恢复的动态变化。6周后，从小鼠获取骨髓（BM）和脾脏样本以检测人细胞。
培养14天后，细胞总数增加了1.78倍。所有接受新鲜和培养的CD34(+)细胞的受体均存活。在骨髓和脾脏中可检测到人细胞、人谱系细胞和人ALU序列。所有移植小鼠的外周血细胞计数均恢复。培养的CD34(+)细胞的植入水平与新鲜CD34(+)细胞相似，但人谱系细胞的百分比高于新鲜CD34(+)细胞。
培养14天的CD34(+)细胞仍保留造血重建能力，并且比新鲜CD34(+)细胞表现出更好的多谱系重建能力。
