异基因间充质干细胞和造血干细胞联合移植治疗难治白血病的初步临床研究

目的　初步探讨异基因间充质干细胞 (MSC)联合造血干细胞移植 (HSCT)治疗难治白血病的安全性、疗效 ,以及对移植物抗宿主病 (GVHD)和造血重建的影响。方法　采用无血清体系培养扩增供者骨髓MSC ,联合移植 3例难治白血病 ,其中 1例联合HLA匹配、2例联合HLA半匹配HSCT。植活证据采用短串联重复序列 聚合酶链反应 (STR PCR)、染色体核型或荧光原位杂交技术(FISH)检测。结果　联合移植无明显不良反应。中性粒细胞≥ 0 5× 10 9/L和血小板≥ 2 0× 10 9/L的时间分别为移植后 13、16、15天和 2 5、30、39天。分别于移植后 16、2 3、2 8天骨髓检查显示完全供者型植入。发生Ⅰ、Ⅲ度急性GVHD各 1例 ,经甲基泼尼松龙治疗后有效控制。 3例患者移植后均获完全缓解 (CR) ,2例持续CR 2 4 8和 15 6天 ,1例移植后 94天复发 ,经供者淋巴细胞输注 (DLI)治疗后再获CR。结论　MSC联合HLA匹配和半匹配HSCT治疗难治白血病安全、有效 ,MSC对异基因造血干细胞移植 (Allo HSCH)造血重建和GVHD的影响有待进一步研究     

异基因外周血造血干细胞移植治疗骨髓增生异常综合征转化的急性髓细胞白血病1例及文献复习

目的探讨异基因外周血造血干细胞移植(强行移植)治疗原发耐药的急性髓细胞白血病(由骨髓增生异常综合征转化)的可行性。方法患者,女,16岁,供者为其胞弟,HLA配型全相合。预处理方案:改良BuCy。移植单个核细胞(MNC)6.22×108/kg(受者),CD34+占6.1%。结果移植后+14d粒系植入,+21巨核系植入。+30d染色体转为46,XY。移植后合并肺炎克雷伯杆菌败血症。无急慢性移植物抗宿主病发生。随访14个月,患者获得无病生存,生活质量良好。结论对于原发耐药的急性髓细胞白血病(由骨髓增生异常综合征转化),如有合适供者,宜尽早行造血干细胞移植。     

两种rhG-CSF制剂用于自体外周造血干细胞移植的效果比较

目的探讨两种重组人粒细胞集落刺激因子吉赛欣(国产rhG-CSF)和惠尔血(进口rhG-CSF)在外周造血干细胞(PBSC)动员及移植后造血恢复中的效能差异。方法选择2000-01～2003-05在本院进行自体外周造血干细胞动员的47例血液肿瘤患者,随机分为两组,在化疗后白细胞降至最低点时分别接受国产rhG-CSF和进口rhG-CSF进行干细胞动员,比较rhG-CSF使用时间及采集所得CD34+细胞数量,其中接受干细胞回输的41例再随机分组,在预处理和干细胞回输后,外周WBC达到0×109/L时分别接受两种rhG-CSF促进造血恢复,比较外周血中性粒细胞绝对计数(ANC)恢复至≥1.5×109/L的天数及rhG-CSF使用时间。结果47例进行干细胞动员的患者中,24例应用进口rhG-CSF,23例应用国产rhG-CS,使用时间分别为6.17±2.64d和5.78±1.83d,所得采集物CD34+细胞总数分别为(5.90±5.06)×106/kg体重和(5.13±6.07)×106/kg体重,两者比较无显著性差异。41例接受自体干细胞回输的患者,22例应用进口rhG-CSF,19例应用国产rhG-CSF,使用时间分别为10.86±2.41d和10.83±4.75d,ANC恢复至≥1.5×109/L的时间分别为9.07±1.50d和10.00±4.20d,两者比较亦无显著性差异。结论两种rhG-CSF制剂在PBSC动员、PBSC回输后造血恢复等方面无明显差异,均可供临床上选择使用。     

自体外周血造血干细胞移植治疗系统性红斑狼疮9例临床疗效探讨

目的 :探讨自体外周血造血干细胞移植 (Autologousperipheralbloodstemcellstransplantation ,ABSCT)治疗系统性红斑狼疮 (SLE)的临床效果。方法 :选择 9例SLE患者 ,应用BaxterCS - 30 0 0plus连续血流式血细胞分离机进行自体外周血造血干细胞采集。预处理方案 :分次全淋巴照射 (TLI) 12 - 16Gy ,环磷酰胺 5 0mg/kg(- 3,- 2 ,- 1d) ,ATG 10mg/kg(+1,2d)。外周血干细胞动员 :环磷酰胺 2 0 0 0mg/m2 ,G -CSF 5ug/kg/d。结果 :造血干细胞采集量 :MNC 3.32× 10 9/Kg(2 .6 7- 4.2 8) ,患者造血功能重建迅速 ,免疫重建特征为CD4 ,CD19细胞减低 ,CD8,CD16+ 56细胞升高。随访 8例完全缓解 ,1例部分缓解 ,未发生移植相关的严重并发症。结论 :ABSCT治疗SLE近期临床疗效显著 ,远期疗效尚须进一步探讨。疗效机理可能与预处理后组织中免疫病理细胞的减少 ,免疫球蛋白降低 ,自身抗体量的下降有关。     

自体外周血干细胞移植支持大剂量化疗治疗精原细胞瘤2例报告

应用自体外周血干细胞移植支持大剂量化疗治疗精原细胞瘤 2例。干细胞动员采用 G- CSF+ VP16或单用 G- CSF,采集循环血量为 5 30 0～ 2 30 0 0 ml,采集 MNC5 .3× 10 9～ 13.39× 10 9,CD34+ 1.95 6× 10 8～ 9.5 4× 10 8。预处理方案为 CVC或 IEP方案 ,2 4小时及 36小时回输干细胞。移植后恢复均顺利 ,现已无病生存 10个月～ 3年。     

自体骨髓和/或外周血干细胞移植治疗14例恶性血液病

为研究自体骨髓和外周血干细胞联合移植(ＢＭＴ/ＰＢＳＣＴ)的疗效、安全性和费用 ,回顾性分析本院 1997年 6月～ 1998年 6月经治的 4例恶性血液病 ,并与以前进行的单一ＢＭＴ或ＰＢＳＣＴ组 10例作比较。资料和方法病例 :3例恶性淋巴瘤 ,1例急粒白血病 ,行自体ＢＭＴ/ＰＢＳＣＴ ;白血病 8例 ,行单一骨髓移植(ＢＭＴ) ;白血病 2例 ,行单一外周血干细胞移植(ＰＢＳＣＴ)。此三组的性别、年龄 (平均 34 9岁 )、预处理方案可比。ＢＭＴ组植入单个核细胞 (ＭＮＣ)数≥ 1 0× 10 8/ｋｇ(受体体重 )。ＰＢＳＣＴ组植入ＭＮＣ≥ 2 0× 10 8/ｋ…     

血液病患者骨髓间充质干细胞的增殖分化特点

目的:研究血液病患者骨髓间充质干细胞(MSC)的体外增殖分化特点.方法:用H4434甲基纤维素半固体培养基培养168例血液病患者骨髓单个核细胞14 d,分析贴壁梭形细胞形态和生长等级特点;CD29、CD44、α-SMA、α-醋酸奈酚酯酶和碱性磷酸酶组化染色鉴定MSC特性和分化特点.结果:所有患者贴壁细胞为梭形,核圆或椭圆形,瑞氏染色胞浆淡蓝,核为紫红色;α-醋酸奈酚酯酶染色阳性.所有患者贴壁细胞的CD29和CD44阳性率在90%～95%之间,碱性磷酸酶和α-SMA阳性率分别为25%和30%.再生障碍性贫血、急性白血病、骨髓异常增生综合征和骨髓增殖性疾病4组患者贴壁细胞生长等级不同,再生障碍性贫血组分别高于其他3组(P<0.05).结论:血液病患者MSC非诱导体外培养广泛表达骨和肌型细胞标志;再生障碍性贫血患者MSC生长等级显著高于急性白血病、骨髓异常增生综合征和骨髓增殖性疾病.     

骨髓间质干细胞移植对心肌梗死后创伤修复的影响

目的:探讨骨髓间质干细胞(MSC)移植对心肌梗死后创伤修复的影响,为临床MSC移植提供理论基础。方法:制备大鼠急性心肌梗死模型,分离培养同种异体MSC。术后第7天,心肌梗死区植入20μL用BrdU标记的MSC[(0.5~1.0)×105/μL]。3周后,免疫组化检测梗死区BrdU,对梗死范围、梗死区心室游离壁厚度和胶原容积分数进行图像分析,并进行统计学分析。结果:MSC移植后3周,梗死区可见大量BrdU染色阳性细胞。处理组梗死面积为(24.91±3.62)%,低于对照组的(40.41±4.55)%;梗死区心室游离壁厚度(2.45±0.39)mm,较对照组(1.65±0.41)mm增加;胶原容积分数(57.11±3.42)%,较对照组的(81.36±5.31)%减小,差别均有统计学意义。结论:MSC移植后可促进心肌梗死后创伤修复过程,改善梗死区结构,具有良好的临床应用价值。     

异基因造血干细胞-间充质干细胞协同移植治疗急性粒细胞白血病(附2例报道)

目的探讨急性粒细胞白血病异基因造血干细胞—间充质干细胞协同移植的效果。方法培养扩增供者骨髓间充质细胞,经形态学及细胞表面分子加以鉴定,并与外周血造血干细胞协同移植给患者。结果输注供者的MSC总数分别为2×106/kg和7.7×106/kg,中性粒细胞>0.5×109/L以及血小板>20×109/L的中位时间分别是12d和14d。例1出现了Ⅰ度aGVHD,Ⅱ度cGVHD,已存活11个月;例2未出现急、慢性GVHD,现已存活6个月余。结论异基因造血干细胞-间充质干细胞协同移植可加速造血功能恢复,对GVHD的发生及严重程度有一定抑制作用,但还需加大病例数进一步观察。     

自体造血干细胞移植治疗晚期恶性实体瘤

目的观察自体造血干细胞 (aulologoushematopoiticstemcelltransplantation ,AHSCT)对晚期恶性实体瘤的疗效及副作用。方法 6例患者中 ,1例行自体骨髓移植 (antologousbonemarrowtranspla tion ,ABMT) ,5例行自体外周血干细胞移植 (autologousperipheralbloodstemcelltransplantationAPBSCT)。预处理方案 :乳腺癌为CEP(CTX ,VP-1 6,DDP) ,恶性淋巴瘤为CEA(CTX ,VP-1 6,Ara-c)方案 ,采集的干细胞 1例 4℃保存 ,72小时内回输 ,其余 5例 - 80℃保存。结果APBSCT动员采集单个核细胞数 7 0 2× 1 0 8 kg ,ABMT和APBSCT后 ,患者中性粒细胞计数恢复到 0 5× 1 0 9/L ,血小板达 50× 1 0 9/L以上的时间平均为 1 0 8天和 1 6 2天。毒副作用主要是骨髓抑制和消化道反应 ,随访至今死亡 2例 ,复发 2例 ,2例无瘤生存。结论AHSCT治疗晚期乳腺癌近期疗效肯定 ,可提高生活质量 ,对恶性淋巴瘤的治疗有效 ,但对反复复发患者需移植后作进一步治疗。     

Reversing breast cancer stem cell into breast somatic stem cell

Stem cells have an important role in cell biology, allowing tissues to be renewed by freshly created cells throughout their lifetime. The specific micro-environment of stem cells is called stem cell niche; this environment influences the development of stem cells from quiescence through stages of differentiation. Recent advance researches have improved the understanding of the cellular and molecular components of the micro-environment--or niche--that regulates stem cells. We point out an important trend to the study of niche activity in breast cancers. Breast cancer has long been known to conserve a heterogeneous population of cells. While the majority of cells that make up tumors are destined to differentiate and eventually stop dividing, only minority populations of cells, termed cancer stem cell, possess extensive self renewal capability. These cancer stem cells possess characteristics of both stem cells and cancer cells. Breast cancer stem cells reversal to breast somatic stem cells offer a new therapy, that not only can stop the spread of breast cancer cells, but also can differentiate breast cancer stem cells into normal breast somatic stem cells. These can replace damaged breast tissue. Nevertheless, the complexity of realizing this therapy approach needs further research.     

Cardiac-derived stem cells

Current scientific dogma holds that cardiomyocyte stem cells do not exist in the adult mammalian heart, and furthermore, that there is little, if any, potential for the regeneration of damaged myocardium. In order to approach this topic, I have begun with a brief overview of advances in stem cell research in other organ systems, such as the bone marrow and the brain. Very recent progress in cardiac stem cell research is then discussed, which indicates that a cardiomyocyte progenitor cell contributes to cardiomyocyte replacement throughout life. This progenitor cell may reside in the heart itself, or derive from a circulating marrow stem cell.     

Glioma gene therapy using induced pluripotent stem cell derived neural stem cells

Using neural stem cells (NSCs) with tumor tropic migratory capacity to deliver therapeutic genes is an attractive strategy in eliminating metastatic or disseminated tumors. While different methods have been developed to isolate or generate NSCs, it has not been assessed whether induced pluripotent stem (iPS) cells, a type of pluripotent stem cells that hold great potential for regenerative medicine, can be used as a source for derivation of NSCs with tumor tropism. In this study, we used a conventional lentivirus transduction method to derive iPS cells from primary mouse embryonic fibroblasts and then generated NSCs from the iPS cells. To investigate whether the iPS cell derived NSCs can be used in the treatment of disseminated brain tumors, the cells were transduced with a baculoviral vector containing the herpes simplex virus thymidine kinase suicide gene and injected into the cerebral hemisphere contralateral to a tumor inoculation site in a mouse intracranial human glioma xenograft model. We observed that NSCs expressing the suicide gene were, in the presence of ganciclovir, effective in inhibiting the growth of the glioma xenografts and prolonging survival of tumor-bearing mice. Our findings provide evidence for the feasibility of using iPS cell derived NSCs as cellular vehicles for targeted anticancer gene therapy.     

造血干细胞衰老机制研究

造血干细胞(hematopoietic stem cell,HSC)具有自我更新功能和生成多个系列免疫造血细胞的多分化功能,终生维持机体造血。证据显示HSC会出现衰老,并与机体老龄化及各种损伤情况下出现造血系统应激能力下降有关。DNA损伤反应在诱导HSC衰老的各种机制中起重要作用。HSC发生衰老的机制的研究将对降低放化疗毒副作用,保护辐射等环境因素的损伤,了解白血病的干细胞起源及治疗逃逸机制提供有价值的科研资料,为药物筛选的靶点提供试验依据。     

Embryonic stem cell therapy

Stem cell therapies, particularly those using embryonic stem cells, offer a novel approach to treating disease. There is an ongoing effort to develop tools and reagents to assist in understanding stem cells at a research level. In addition to these research tools, making stem cell therapy a reality requires the development of tools that enable the translation of research into viable therapies. Three sets of tools are discussed in this article: tools enabling stem cell scale-up and manufacture to GMP standards, tools addressing the behavior of cells in animal models, and tools to assess transplanted cells in early clinical trials. The development of such tools will address many of the safety and efficacy questions that are likely to arise as stem cell therapies move from bench to bedside.