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1.
外周血干细胞的动员和采集及移植后造血重建研究   总被引:24,自引:1,他引:24  
对24例外周血干细胞(PBSC)移植的三种动员方案、采集及移植后的PBSC数量与造血重建关系进行研究。24例中急性白血病13例,多发性骨髓瘤6例,非霍奇金淋巴瘤4例,晚期乳癌1例。三组动员方案:化疗+四氢叶酸+氟美松;化疗+rhGM-CSF+氟美松;化疗+rhG-CSF+氟美松。用流式细胞仪双染色直接免疫荧光法测定rhG-CSF组中9例CD34+CD33+细胞。结果表明:rhG-CSF组PBSC产率最高,平均每例单个核细胞(9.86±6.01)×108/kg,CFU-GM(21.08±16.86)×104/kg,CD34+细胞数与CFU-GM正相关(r=0.68)。动员后再用rhG-CSF1周,CD34+细胞显著增多,可连续每日采集,当≥5×106/kg即可停止;采集与回输的PBSC数与造血重建时间密切相关。  相似文献   

2.
用两种单克隆抗体(单抗)标记脐血造血祖细胞表面抗原(HPCA,CD34)用流式细胞仪分析,并比较两种单抗标记的CD34^+细胞与体外培养的粒单细胞集落形成单位(CFU-GM),红系爆式集落形成单位(BFU-E)和混合集落形成单位的相关性,结果表明脐血有核细胞中,抗HPCA-2FITC阳性的细胞占1.05±0.72%,Tuk3(纯抗体)阳性细胞占2.06±1.25%,差别显(P<0.05),每μl  相似文献   

3.
造血生长因子对脐血CD34^+细胞的体外扩增作用   总被引:21,自引:2,他引:21  
为探讨造血生长因子不同组合方式对脐血CD34+细胞的扩增作用、扩增细胞性能改变及合理的扩增时机,利用吸附单克隆抗体——磁珠分离系统分选出95%~99%纯度的脐血CD34+细胞,在体外液体培养体系中,经重组人干细胞因子(rhSCF)、白细胞介素(IL)3、IL-6、粒-巨噬细胞集落刺激因子、粒细胞集落刺激因子(G-CSF)和红细胞生成素(Epo)的不同组合扩增1~3周。结果表明,脐血CD34+细胞具有良好的扩增效应,细胞总数、集落形成细胞(CFCs)和CD34+细胞可分别扩增500.0±82.5倍、94.0±28.6倍和24.0±3.8倍;SCF是扩增最重要的生长因子,SCF+IL-3+IL-6+G-CSF+Epo组合的扩增效率最高;CFCs和CD34+细胞在1~2周维持在较高水平,以后即快速衰减;此外,扩增也加速了细胞的分化,至第3周时93%CFCs为CFU-GM,极少部分形成BFU-E和CFU-GEMM。因此,合理组合生长因子,把握适宜扩增时机,将有助于扩增细胞在质和量上满足移植和基因治疗等的要求。  相似文献   

4.
为估算经粒细胞集落刺激因子(G-CSF)动员的外周血中祖细胞和多能因干细胞的含量,将不同数量的经G-CSF动员的外周血单个核细胞(PBMC)移植到经全身照射(TBI)的同系小鼠,与骨髓移植(BMT)组比较受体鼠外周白血细胞恢复动态及长期存活率。结果显示,输入相同剂量(1×10^6)细胞14和21天时,经动员的PBMC移植鼠的白细胞数远较BMT鼠为高(P〈0.05),说明前者含有更多的祖细胞。150  相似文献   

5.
为深入研究造血干细胞(HSC)的生物学功能,根据表面标志,采用流式细胞仪分离和富集了小鼠骨髓中两类不同特征的HSC亚群c-kit+Sca-1+和c-kit+Sca-1-细胞。体外集落培养表明c-kit+Sca-1+细胞对IL-3或GM-CSF单独刺激无反应,而c-kit+Sca-1-细胞则可形成大小不等的细胞集落。多种造血生长因子[IL-3+IL-6+GM-CSF+G-CSF+Epo+干细胞生长因子(SCF)]联合可诱导c-kit+Sca-1+细胞形成大的细胞集落,但在培养12天时尚无BFU-E和粒、单核、红和巨核系细胞集落(CFU-GEMM)形成;而同样条件下,c-kit+Sca-1-细胞可形成BFU-E和CFU-GEMM以及其它各类集落。体内实验表明c-kit+Sca-1+细胞在培养8天时无脾集落(CFU-S)形成,与c-kit+Sca-1-细胞显著不同(P<0.01)。相反,c-kit+Sca-1+细胞包含了骨髓造血重建活性细胞,而c-kit+Sca-1-细胞则缺乏。结果提示c-kit+Sca-1+细胞为早期的具重建长期造血功能的干细胞,而c-kit+Sca-1-细胞则为相对分化后期的造血祖细胞。  相似文献   

6.
G—CSF对化疗后外周血干细胞动员作用的影响   总被引:1,自引:0,他引:1  
通过对11例急性白血病患单独化疗与化疗后加用粒细胞集落刺激因子(G-CSF)的对比,动态观察了G-CSF对外周血造血干细胞(PBSC)的动员作用。发现化疗后加用G-CSF比单用化疗的粒-巨噬细胞集落形成单位(CFU-GM)增加5.1倍,红系式集落形成单位(BFU-E)增加4.5倍。G-CSF还可使CFU>100/ml和BFU-E>200/ml的持续时间延长;化疗后CFU-GM的最高值提早出现,而  相似文献   

7.
G-CSF对化疗后外周血干细胞动员作用的影响   总被引:1,自引:0,他引:1  
通过对11例急性白血病患者单独化疗与化疗后加用粒细胞集落刺激因子(G-CSF)的对比,动态观察了G-CSF对外周血造血干细胞(PBSC)的动员作用。发现化疗后加用G-CSF比单用化疗的粒-巨噬细胞集落形成单位(CFU-GM)增加5.1倍,红系爆式集落形成单位(BFU-E)增加4.5倍。G-CSF还可使CFU-GM>100/ml和BFU-E>200/ml的持续时间延长;化疗后CFU-GM的最高值提早出现,而不影响BFU-E/CFU-GM比值。结果表明,化疗后加用G-CSF可明显提高PBSC的收集效率,G-CSF是一种有效的PBSC动员剂。  相似文献   

8.
用两种单克隆抗体(单抗)标记脐血造血祖细胞表面抗原(HPCA,CD_(34))用流式细胞仪分析,并比较两种单抗标记的细胞与体外培养的粒单细胞集落形成单位(CFU-GM),红系爆式集落形成单位(BFU-E)和混合集落形成单位的相关性,结果表明脐血有核细胞中,抗HPCA-2-FITC阳性的细胞占1.0510.72%,Tuk3(纯抗体)阳性细胞占2.06±1.25%,差别显著(P<0.05),每μl脐血两种单抗标记的细胞分别为96.56±56.64和231.40±163.93(P<0.05),变异系数依次为58.47%和68.43%。尽管抗HPCA-2-FITC阳性细胞与阳性细胞数量呈显著正相关,但前者与CFU-GM,BFU-E,CFU-Mix以及CFUs(CFU-GM+BFU-E+CFU-Mix)均呈正相关,而后者仅与CFU-GM,CFUs呈正相关。研究结果提示在检测造血祖细胞时,用抗HPCA-2-FITC代替可降低假阳性,获得较好的细胞与CFU间的线性关系。  相似文献   

9.
单个CD34^+CD38^+和CD34^+CD38^—l细胞的增殖与分化性能   总被引:3,自引:0,他引:3  
为深入探讨CD34^+细胞亚群的不均一性及单个细胞的增殖分化特性,利用FACS Vantage自动细胞分选系统(ACDU)分选出单个CD34^+亚群细胞:CD34^+CD38^+和CD34^+CD38^-,在96孔板无血清培养体系中,经SCF、IL-3、IL-6、GM-CSF、G-CSF和Epo刺激,14-18天后扩增形成原代集藩。单个CD34^+CD38^+细胞形成原代集落的比例为38.0%±9  相似文献   

10.
为估算经粒细胞集落刺激因子(G-CSF)动员的外周血中祖细胞和多能造血干细胞的含量,将不同数量的经G-CSF动员的外周血单个核细胞(PBMC)移植到经全身照射(TBI)的同系小鼠,与骨髓移植(BMT)组比较受体鼠外周血白细胞恢复动态及长期存活率。结果显示,输入相同剂量(1×106)细胞14和21天时,经动员的PBMC移植鼠的白细胞数远较BMT鼠为高(P<0.05),说明前者含有更多的祖细胞。150天存活期的观察发现,移植≥1×106经动员的PBMC或≥1×105骨髓细胞均可使80%左右的受体鼠长期存活;而移植1×105经动员的PBMC或1×104骨髓细胞均使50%受体鼠长期存活,提示经动员的外周血中多能造血干细胞的含量约为同样数目骨髓细胞的1/10。  相似文献   

11.
目的:分析恶性血液系统疾病患者外周血造血干细胞动员与采集过程中的影响因素。方法:对50例血液系统恶性疾病患者在东南大学附属中大医院血液科进行外周血造血干细胞动员。对患者年龄、性别、动员方案、疾病状态、采集机器等因素进行分析,评估以上因素对干细胞动员结果的影响,并分析了采集前白细胞、血红蛋白、血小板的数量与采集的CD34^+细胞计数的相关性。结果:动员方案对CD34^+细胞采集数及CD34^+细胞采集成功率的影响有显著性影响,而性别、年龄、确诊到动员间隔时间、既往化疗方案、骨髓受累与否等对干细胞采集数量影响并不显著。采集前外周血白细胞数量及血红蛋白数量与采集的CD34^+细胞数呈正相关。采集前外周血中白细胞计数及单个核细胞计数与采集成功密切相关。结论:化疗联合细胞因子的动员方案采集造血干细胞优于单用细胞因子的动员方案。通过采集前白细胞计数及单个核细胞计数确定合适的采集时机,可以提高采集的成功率。  相似文献   

12.
To improve the selection of donors for allogeneic stem cell transplantation, it is important to identify reliable parameters that predict CD34+-cell yields after granulocyte-colony stimulating factor (G-CSF)-induced peripheral blood stem cell (PBSC) mobilization. We retrospectively investigated the peripheral blood (PB) kinetics of white blood cells (WBCs), CD34+ cells, matrix metalloproteinases (MMP)-9 and -2, and tissue inhibitors of metalloproteinases (TIMP)-1 and -2 in 15 healthy donors during their treatment with G-CSF. All donors received 10 microg/kg of recombinant human G-CSF once a day subcutaneously. Leukapheresis was initiated after 4 days of G-CSF treatment, and G-CSF treatment continued until the last day of leukapheresis. WBC and CD34+ cell numbers in the PB rose after 2 and 3 or 4 days of G-CSF treatment, respectively. The PB CD34+ cell numbers on day 4 correlated weakly with the increase in WBC counts from day 1 to day 2 (R(2) = 0.254, P = 0.056). There were also positive correlations between the CD34+ cell numbers in the PBSC products on day 4 and the CD34+ cells in the PB on days 1 and 4 (R(2) = 0.768, P < 0.0001 and R(2) = 0.816, P < 0.0005, respectively). The MMP-9 plasma levels on days 1 and 4 also correlated positively with the day 4 circulating CD34+ cell numbers (R(2) = 0.393, P < 0.05 and R(2) = 0.406, P = 0.01, respectively). In conclusion, the CD34+ cell numbers in the PB steady state may be a useful parameter selecting allogeneic PBSC donors.  相似文献   

13.
BACKGROUND: Allogeneic transplantation of granulocyte-colony- stimulating factor (G-CSF)-mobilized peripheral blood progenitor cells (PBPCs) from normal related donors is effective in achieving engraftment with a relatively short period of posttransplantation aplasia. The optimal dose and composition of PBPC transplants are unknown. The CD34+/Thy-1dim progenitor cell subset is enriched for putative stem cells. STUDY DESIGN AND METHODS: The kinetics of the primitive subpopulation were prospectively studied in nine normal donors receiving recombinant human G-CSF (6 microg/kg) subcutaneously twice daily for 6 days for collection of PBPCs for allogeneic transplantation. RESULTS: The concentration (mean +/− SD) of the circulating CD34+/Thy-1dim subset increased from a baseline of 0.9 +/− 0.9 × 10(3) to 29.2 +/− 22.1 × 10(3) per mL on Day 4 and 38.0 +/− 29.8 × 10(3) per mL on Day 6. The level of CD34+/Thy-1dim cells was closely correlated with the overall level of CD34+ cells. At baseline, CD34+/Thy-1dim cells composed 21.1 percent of the total CD34+ cells, increasing to 36.3 percent at the peak of mobilization. CONCLUSION: CD34+/Thy-1dim cells are optimally mobilized on Days 4 to 6 of recombinant human G-CSF treatment.  相似文献   

14.
目的:探讨急性白血病患者外周血干细胞动员、采集的效率及影响因素。方法对37例急性白血病患者经化疗+生长因子动员,动员第5~10天,当外周血WBC>5×109/L,CD34+>20个/μl时,使用CS-3000 Plus血细胞分离机(Baxter公司)进行外周血干细胞采集;采集前外周血WBC分类单个核细胞(MNC,包括幼稚细胞、淋巴细胞及单核细胞),计算MNC%×WBC计数>(4~6)×109/L预计需要采集循环血容量。分析不同化疗动员时机、疾病、年龄、性别的动员采集,并对采集前外周血进行白细胞计数、分类以及干细胞采集物进行WBC计数、分类和CD34+检测。结果所有患者均成功动员和采集到了外周血干细胞(MNC>6×108/kg,CD34+>2×106/kg),并成功造血重建。急性髓细胞白血病患者采集所获得的 MNC 与急性淋巴细胞白血病患者采集获得无明显差异;但急性淋巴细胞白血病患者采集获得CD34+细胞明显较多(P=0.015);动员时机的化疗病程与采集物的CD34+细胞呈负相关,动员时机≤3个疗程与≥6个疗程比较,前者CD34+细胞明显较好,具有统计学差异(P=0.028);外周血MNC数值与采集物MNC及CD34+细胞具有相关性(r=0.600,P=0.00;r=0.510,P=0.001)。结论根据不同的疾病、性别、年龄,采用不同的动员采集方案,一般可以成功采集。外周血 MNC 数值对采集物中 CD34+细胞的总量具有一定预测意义。  相似文献   

15.
At the moment, PBSC collections can be performed using semi-automated or automated cell separator devices. The automated methods offer the advantages of a decreased working load for dedicated personnel and high standardization of the collection procedure. Herein we report our single institutional experience in 80 PBSC collections employing the new automated COM.TEC Fresenius autoMNC program that provides the ability to predict the total number of CD34(+) cells collected, based on the pre-leukapheresis CD34(+) cell count in peripheral blood. Fourty-eight patients and 21 healthy donors were mobilized with chemotherapy + G-CSF or G-CSF alone, respectively. Eighty leukapheresis collections were performed starting with a CD34(+) cell count in peripheral blood at least of 20/microL. Collection parameters and related side effects were evaluated. The mean CD34(+) cell collection efficiency in patients and donors was 81.8% (sd 27.6) and 95.1% (sd 15.6), respectively. The mean difference between real and predicted CD34(+) cells was +30.2% (sd 92.9) for patients and +4.6% (sd 30.3) for donors. The mean leukapheresis bag volume was 240.7 ml (sd 67.3) and 310.3 ml (sd 86.8) with a mean HCT of 10.9% (sd 34.4) and 9.2% (sd 3.9) for patients and donors, respectively. The automated PBSC collection with the new program COM.TEC Fresenius autoMNC demonstrated a very high CD34(+) cell collection efficiency. Moreover, the ability to predict the CD34(+) cell yield permits improved management of the leukapheresis collection, with the only disadvantage of larger leukapheresis volume and higher hematocrit.  相似文献   

16.
Peripheral blood stem cells (PBSC) have nearly replaced bone marrow (BM) as the preferred source of hematopoietic rescue for patients undergoing high-dose chemotherapy. However, some patients fail to mobilize sufficient numbers of PBSC into the peripheral blood thereby putting high-dose chemotherapy at risk. The present article reviews mobilization of PBSC with a special focus on poor mobilizers. Under steady-state conditions less than 0.05% of the white blood cells (WBC) are CD34+ cells. Chemotherapy results in a 5-15-fold increase of PBSC. Combining chemotherapy and growth factors increases CD34+ cells up to 6% of WBC. Several factors affect the mobilization of PBSC: age, gender, type of growth factor, dose of the growth factor and in the autologous setting patient's diagnosis, chemotherapy regimen and number of previous chemotherapy cycles or radiation. Poor mobilizers are defined as patients with less than 10 CD34+ cells/mul in the peripheral blood during mobilization. Promising approaches for those patients rely on remobilization, use of high doses of granulocyte-colony stimulating factor (G-CSF), or the combination of G-CSF and granulocyte macrophage (GM)-CSF, which successfully mobilized the majority of poor mobilizing patients. New agents such as long lasting variants of G-CSF and CXCR4 antagonists are at the horizon and studied in clinical trials as mobilizing agents. Muscle and bone pain are frequent adverse events in stem cell mobilization but are usually tolerated under the use of analgesics. Large volume apheresis (LVL) with a processed volume of more than 4-fold patient's blood volume is an approach to increase the CD34+ yield in patients with low CD34+ pre-counts resulting in higher yields of CD34+ cells for transplantation. Processing of more blood in LVL is achieved by an increase of the blood flow rate and an altered anticoagulation regimen with the occurrence of more citrate reactions.  相似文献   

17.
G-CSF的不同方案动员正常供者外周血干细胞的效果比较   总被引:1,自引:0,他引:1  
为了研究粒细胞集落刺激因子(rhG-CSF)动员的最佳动员方案,对HLA完全相合的非清髓异基因外周造血干细胞移植的供者60例进行回顾性分析。结果显示:供者rhG-CSF10μg/(kg·d)的动员方案组所采集的单个核细胞及CD34+细胞数明显高于供者rhG-CSF5μg/(kg·d)的动员方案组(P<0.05)。供者rhG-CSF10μg/(kg·d)的动员方案在第4天或第5天采集时所获得的单个核细胞及CD34+细胞数无统计学差别。供者不同rhG-CSF动员剂量及采集时间所获得的CD3+、CD4+、CD8+细胞的百分比无统计学差别。结论:供者10μg/(kg·d)的动员效果明显优于供者5μg/(kg·d)的动员效果。供者10μg/(kg·d)的动员方案在第4天或第5天采集时效果无统计学差异,但第4天采集可缩短动员天数,降低费用,因此对供者采用10μg/(kg·d)动员方案时在第4天采集单个核细胞效果更好。  相似文献   

18.
Failure of autologous peripheral blood CD34+ stem cells collection can adversely affect the treatment modality for patients with hematological and nonhematological malignant diseases where high dose chemotherapy followed by hematopoietic stem cell transplantation has become part of their treatment. Plerixafor in conjunction with G-CSF is approved for clinical use as a mobilization agent. The clinical efficacy of Plerixafor in CD34+ cells collection was analyzed in our institution. A total of 13 patients aged 1–15,5 years received Plerixafor in combination with G-CSF: 7 with neuroblastoma, 2 with Ewing’s sarcoma and single patients with Hodgkin’s lymphoma, germ cell tumor, retinoblastoma and Wilms tumor. Twelve patients (923%) achieved CD34+ cell counts of ≥ 20 × 106/L after 1–7 doses of Plerixafor. The average 9,9 - fold increase in number of CD34+ cells were achieved following the first dose and 429 - fold after second dose of plerixafor. Among the 13 patients, 12 yielded the minimum required cell collection of 2 × 106/kg within an average of 2 doses of Plerixafor. The mean number of apheresis days was 1.75. The median total number of collected CD34+ cells was 982 × 106/kg. Plerixafor enables rapid and effective mobilization, and collection of sufficient number of CD34+ cells.  相似文献   

19.
The role of leukocyte-endothelial cell interactions during granulocyte colony-stimulating factor (G-CSF)-induced stem cell mobilization is unclear. To examine endothelial activation during this process, we determined levels of circulating endothelial adhesion molecules in healthy donors undergoing G-CSF-mobilized stem cell collection. Plasma levels of soluble (s) E-selectin, soluble intercellular adhesion molecule-1 (sICAM-1), and soluble vascular cell adhesion molecule-1 (sVCAM-1) were serially determined by enzyme-linked immunosorbent assays in 10 healthy donors during G-CSF-stimulated stem cell mobilization. There was a significant increase in plasma levels of all three endothelial adhesion molecules (sE-selectin, p = 0.01; sICAM-1, p = 0.003; sVCAM-1, p = 0.0002) between day 1 and day 5 of G-CSF stimulation, but only sVCAM-1 concentrations exceeded the range obtained from unstimulated controls in all stem cell donors. Increases of sCAM were accompanied by increased numbers of white blood cells and CD34(+) progenitors in peripheral blood. G-CSF-stimulated peripheral blood progenitor cells (PBPC) mobilization results in increased levels of circulating endothelial adhesion molecules that were most evident for VCAM-1 molecules. Because soluble VCAM-1 remains active in binding to the VLA-4 receptor on CD34(+) cells, it may reduce stem cell adhesiveness to endothelial cells and to bone marrow microenvironment.  相似文献   

20.
目的:研究粒细胞集落刺激因子(G-CSF)联合普乐沙福对异基因造血干细胞移植(allo-HSCT)的亲缘健康供者外周血造血干细胞动员的效果及安全性。方法:回顾性分析2019年4月至2021年4月在河北燕达陆道培医院采用G-CSF联合普乐沙福动员方案的亲缘健康供者33例(观察组),应用G-CSF细胞动员d 4采集骨髓,d 5采集外周血造血干细胞(PBSC),d 5晚加用普乐沙福,并于d 6再次采集PBSC。随机选取历史同期采用单独G-CSF方案动员的亲缘健康供者46例作为对照组,分析2组供者d 5和d 6 PBSC采集物中CD34+细胞计数。以调查问卷的方式观察供者普乐沙福给药后的不良反应。分析接受"G-CSF+普乐沙福"动员方案的allo-HSCT患者和仅接受"G-CSF"动员方案的造血干细胞移植患者在移植后100天总a GVHD、Ⅲ-Ⅳ度a GVHD、CMV血症和EBV血症的发生方面的差异。结果:观察组在d 5和d 6 PBSC采集物中CD34+细胞数(M±Q)分别为(1.71±1.02)×106/kg和(4.23±2...  相似文献   

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