Effective factors for quantity and quality of auto-peripheral blood stem cells in hematopoietic system of patients with malignant tumor / 中国组织工程研究

ABSTRACT

BACKGROUND:Hematopoietic reconstruction of malignant tumor in hematopoietic system is related to disease itself,pretreatment program and therapeutic tool after transplantation;especially,mobilization.collection and cryopreservation of auto-peripheral blood stem cell play a key role in successful reconstruction of hematopoietic system after transplantation.OBJECTIVE:To investigate the reconstruction of hematopoietic system through mobilization, collection and cryopreservation of auto-peripheral blood stem cell in patients with malignant tumor and analyze the effective factors on quantity and quality of auto-peripheral blood stem cell.DESIGN:Case analysis based on malignant tumor in hematopoietic system.SETTING:Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA;Department of Blood,Zhujiang Hospital,Nanfang Medical University.PARTICIPANTS:A total of 18 patients with malignant tumor in hematopoietic system were selected from Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.Their ages ranged from 1 6to 56 years.Among them,2 patients had acute myelogenous leukemia(AML),1 acute lymphoblastic Ieukemia(ALL),2 lymphoblastic Ieukemia (LL),2 chronic granulocytic leukemia(CGL),4 multiple myeloma(MM),and 7 non.Hodgkin lymphoma.Granulocyte colony-stimulating factor(G-CSF)was made by Chugai Pharmaceutical Company Limited (batch numberN3G31).METHODS:①All patients were mobilized with associated chemotherapy+G-CSF.Associate chemotherapyPatients with leukemia were given 2 g/m2 arabinosyl cytosine every 12 hours from lhe first to the third days and 200 mg/m2 etoposide or 50 mg/m2 fludarabine from the first to the fifth days. In addition patients with MM were treated with arabinosyl cytosine as the same way mentioned above and with 1 g/m2 cyclophosphamide from the first to the second days. And patients with lymphoma were given 2 g/m2 cyclophosphamide from the first to the second days. When numbers of leucocyte of all patients decreased below 1.0×109L-1 after chemotherapy.G-CSF started mobilization and the collection was stopped with 5μg/(kg·d)subcutaneous injection.②When numbers of leucocyte increased to (4.0-10.0)×109 L-1,hemopoietic stem cells of peripheral blood were collected till the amount of mononuclear cells≥4.0×108/kq or numbers of CD34+ cells≥2.0×108/kg.And then,the samples were dealt with cooling device.maintained in liquid nitrogen at-196℃ and defrosted in water bath at 37-40℃.③Focal sites of patients were pretreated with local irradiation with 200 cGy/time and 5 times/week for 4 successive weeks.The total dosage was 40 Gy.At 48 hours later,(55.3±28.7)mL hemopoietic stem cells of peripheral blood were transfused back. And the duration from transfusion to collection was about(56.5±22.3)days.300 μg/d G-CSF was subcutaneously injected into all patients at 1 day after transplantation and the reaction was stopped at the phase of neutrophil≥0.5×109L-1. Finally. Refusing-staining rate of trypan blue of peripheral blood stem cell, amount of mononuclear cells, number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were detected before and after thaw.MAIN OUTCOME MEASURES:①Collection of auto-peripheral blood stem cell;②survival rate and related markers of auto-peripheral blood stem cell after cryopreservation;③hematopoietic reconstruction of auto-peripheral blood stem cell after transplantation.RESULTS:All 18 patients with malignant tumor in hematopoietic system were involved in the final analysis.The mean collection time of auto-peripheral blood stern cell was 12.6 days after chemotherapy.the collection times were 1.9.total number of leucocyte was(8.93±1.27)×1 0.L-1 on the first day,and collection rate of mononuclear cell was (138.33±28.61)%. ②Refusing-staining rate of trypan blue of auto-peripheral blood stem cell was similar before and after cryopreservation[(96.26±1.33)%, (92.75±2.04)%,P＞0.05].in addition,after cryopreservation,recovery rates of mononuclear cells,CD34+ cells and granulation-monophyly progenitor cell were(91.96±1.37)%, (85.94±0.64)%and (87.69±4.53)%,respectively.Collection rate of mononuclear cells,number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were lower in patients with myeloma than in those with leukemia and lymphoma (t=2.524-3.268.P＜0.05).③At 15 days after transplantation,15 patients had the neutrophil≥0.5× 109L-1;at 20 days after transplantation,blood platelet was≥20 × 100 L-1.granulation-monophyly progenitor cells[(18.67-26.82)× 105/kg] of 5 patients grew poorly if the course of chemotherapy was more than 10 times.Among them,3 patients had delayed hematopoietic reconstruction after transplantation of auto-peripheral blood stem cell.CONCLUSION:①High-dose chemotherapy combined with G-CSF can shorten collection time of peripheral blood stem cell and improve collection rate of mononuclear cells.②Increase of chemotherapy times before transplantation can affect quantity and quality of auto-peripheral blood stem cell and cause delayed hematopoietic reconstruction.