摘要
目的采集单倍相合异基因供者外周血和骨髓中的造血干细胞,分析不同采集方式的差异,并进行移植后的效果评价。方法单倍相合异基因造血干细胞移植的受者为2006年6月-2008年12月在第三军医大学新桥医院血液科就诊的56例白血病患者,健康供者为患者的父母或兄弟姐妹。在供者皮下注射重组人粒细胞集落刺激因子进行造血干细胞动员,当外周血单个核细胞(MNC)>4.0×109/L时用血细胞分离机采集外周血中的造血干细胞,采集后1h内输给受者。56例供者根据骨髓造血干细胞的采集方法不同分为3组:第1组(n=5)采用注射器针头半密闭式过滤法,第2组(n=6)采用Thomas不锈钢开放式过滤法,第3组(n=45)采用本课题组的改进方法(用过滤输血器替换注射器针头及不锈钢网)。如若供、受者ABO血型不合,则对采集后的骨髓液进行处理:主侧不合用羟乙基淀粉沉降法去除红细胞,次侧不合采用低温低速离心去除血浆。骨髓液干细胞采集后4h内输给受者。结果56例供者外周血采集量的中位值为56ml,MNC中位值5.15×108/kg,CD34+细胞中位值5.80×106/kg;骨髓液采集量中位值950ml,有核细胞(NC)中位值10.31×108/kg,CD34+细胞中位值11.33×106/kg;外周血和骨髓采集的CD34+细胞总数中位值为13.42×106/kg。3种骨髓干细胞采集方法收集的有核细胞及CD34+细胞的质量无明显差异。供受者ABO血型匹配情况:15对ABO血型主侧不合,17对ABO血型次侧不合,均成功留取有效造血干细胞。移植后21d,2例受者死于颅内出血,余54例移植后30d成为完全供者型植入,且均成功造血重建。结论采用单倍相合异基因供者的外周血和骨髓造血干细胞进行移植,可使白血病患者达到有效的造血重建。采用过滤输血器收集供者骨髓干细胞的方法安全、可行,经此法采集的骨髓有核细胞及CD34+细胞的质量与传统方法无明显差异。
Objective To evaluate the therapeutic effects of transplantation of haploidentical allogeneic stem cells, and the difference between those collected from peripheral blood and from bone marrow. Methods Fifty-six patients with leukemia, admitted to Department of Hematology of Xinqiao Hospital from Jun. 2006 to Dec. 2008, were enrolled as recipients who received transplantation of haploidentical allogeneic stem cells, with their family members including, parents, brothers or sisters as the donors. All donors were given subcutaneous injection of granulocyte colony stimulating factor （G-CSF） for stem cells mobilization. Subsequently, stem cells in peripheral blood were isolated and collected with CS-3000 plus blood cell separation machine when the number of mononuclear cells （MNC） reached over 4.0×109 per liter, and they wer given to recipients within an hour. According to different collection methods in collecting stem cells from bone marrow, 56 donors were divided into 3 groups： bone marrow of Group 1 （n=5） was filtered with Lu Dao-pei＇s method （filter bone marrow with an injection needle）, that of Group 2 （n=6） was filtered with Thomas＇ method （filter bone marrow with stainless steel strainer）, and that of Group 3 （n=43） was filtered with the improved method （syringe needle and stainless steel strainer were replaced with transfusion set with stainless steel mesh）. The erythrocytes of bone marrow should be removed by hydroxyethyl starch （HES） precipitation when major ABO incompatibility was found, while the plasma should be removed when there was minor ABO incompatibility. The stem cells were isolated from bone marrow and infused to recipients within 4 hours after collection. Results The median volume of collected peripheral blood was 56ml, the median amount of MNCs was 5.15×108/kg, and the median amount of CD34＋ cells was 5.80×106/kg, and the median volume of 950ml of bone marrow, including a median number of 10.31×108/kg of nucleated cells and a median number of 11.33×106/kg of CD34＋ cells, were collected from 56 donors. A total amount of CD34＋ cells reached to 13.42×106/kg as obtained from peripheral blood and bone marrow. The quality of nucleated cells and CD34＋ cells showed no significant difference as isolated by the 3 different methods. Major ABO imcompatibility was found in 15 cases and minor ABO imcompatibility in 17 cases, while the stem cells were effectively isolated from them. Among 56 recipients received stem cell transplantation, 2 died of intracranial hemorrhage 21 days after transplantation, and in 54 patients full donor chimerism developed on day 30 after transplantation and achieved complete hematopoietic reconstitution. Conclusions Transplantation of haploidentical allogeneic stem cells, collected from peripheral blood and bone marrow, may lead to a complete hematopoietic reconstitution in the patient with leukemia. The transfusion apparatus with filter as used in the present study for stem cell collection is feasible and effective, with which the quality of nucleated cells and CD34＋ cells isolated is the same compared with that isolated with traditional methods.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2010年第5期584-587,共4页
Medical Journal of Chinese People's Liberation Army
关键词
造血干细胞移植
单倍性
造血干细胞动员
hematopoietic stem cell transplantation
haploidy
hematopoietic stem cell mobilization
