具骨髓间充质干细胞表型的成体干细胞移植造血  

Transplantation of Mesenchymal Derived Stem Cells Followed by G-CSF Injection Can Reconstitute Hematopoiesis of Lethally Irradiated BALB/c Mice

作　　者：呼莹[1] 马丽[1] 马冠杰[1] 姜学英[1] 赵春华[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《中国医学科学院学报》2002年第1期20-24,共5页Acta Academiae Medicinae Sinicae

基　　金：国家攀登计划(95-专-10)资助~~

摘　　要：目的为了探索骨髓成体干细胞是否能重建超致死剂量射线照射后小鼠的造血功能。方法采用液体培养分离，培养，扩增雄性小鼠骨髓成体干细胞，作为供体细胞，将受体雌性小鼠预先经Ｃｓ　　　　１３７全身照射８５０ｒａｄ后在４ｈ内从尾静脉输入细胞。受体小鼠分为５组，每组１０只：ａ正常小鼠不经任何处理；ｂ输入６×１０６ｃｅｌｌ／只（０．３ｍｌ）新鲜全骨髓；ｃ输入６×１０６ｃｅｌｌ／只（０．３ｍｌ）去基质细胞的造血细胞；ｄ输入６×１０６ｃｅｌｌ／只（０．３ｍｌ）经培养后的成体干细胞，输注细胞后第１天上、下午肌肉注射各一次Ｇｒａｎｕｌｏｃｙｔｅ　ｃｏｌｏｎｙ　ｓｔｉｍｕｌａｔｉｎｇ　ｆａｃｔｏｒ（Ｇ－ＣＳＦ）等细胞因子；ｅ注射Ｄ－Ｈａｎｋｓ液０．３ｍｌ　／只。观察各组小鼠的存活情况，小鼠在输注前查尾静脉血，计细胞总数、涂片、分类，输细胞后每周各组分别取小鼠肝、脾、股骨，固定作病理切片，另外一侧股骨冲出骨髓，计数一条股骨有核细胞数、涂片、测定ｃｏｌｏｎｙ　ｆｏｒｍｉｎｇ　ｕｎｉｔ－ｇｒａｎｕｌｏｃｙｔｅ／ｍａｃｒｏｐｈａｇｅ（ＧＭ－ＣＦＵ）值。结果输全骨髓组外周血有核细胞和骨髓有核细胞数及ＧＭ－ＣＦＵ值在输注后的第１，２周都高于单输成?Objective To explore the hematopoietic reconstitution potential of mesenchymal derived stem like cells.Methods We transplanted bone marrow mesenchymal derived stem like cells into lethally irra-diated BALB/c mice.Hematopoietic cells were derived from the non-adherent bone marrow cells24hours after initial culture while murine mesenchymal derived stem like cells from bone marrow of donor mice were cultured for10days before the transplantation.Results All mice of group1and3died in7～8days post irradiation following transplantation,while all the mice from group2and4survived.The time course of hematopoietic reconstitution was then observed.The peripheral blood and bone marrow cell count recovered in the MSC+G-CSF transplanted group and the BM transplanted group after3weeks.Interestingly,CFU-GM number in the MSC+G-CSF transplanted group increased significantly after2weeks and even more than that in the BM transplanted group after3weeks while as CFU-GM colony dropped2weeks after in the BM transplanted group.Spleen colony(CFU-S)number and size of the MSC+G-CSF transplanted group was significantly greater than the BM transplanted group.Furthermore,PCR analysis was performed using peripheral blood cells to determine if any male-derived cells were present.No male-derived cells were found in any of the mice from group1and3.Y-chromosome -specific src gene was found to be dominant in the MSC+G-CSF transplanted group and the BM transplanted group by week4post transplant ation.In addition,we demonstrated that induction with G-CSF lead to CFU-GM colony formation from MSC com partment in vitro.Conclusion These results indicate that under stimulation of G-CSF,mesenchymal derived stem like cells might differentiate into hematopoietic primitive stem cells in vivo and have the capacity to re-establish hematopoiesis in lethally irradiated mice.This study should provide an alternative transplantation tre atment for malignancy.

关 键 词：骨髓 成体干细胞移植 重建造血 间充质干细胞 

分 类 号：R457.7[医药卫生—治疗学] Q26[医药卫生—临床医学]