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体外高效活化扩增人脐带血来源自然杀伤细胞的两种体系的比较研究
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作者 王媞尔 孙运艳 +2 位作者 韩忠朝 张磊升 史明霞 《中国实验血液学杂志》 CAS CSCD 北大核心 2023年第2期553-561,共9页
目的:探讨人脐带血来源的单个核细胞(hUC-MNC),经两种体外培养体系活化扩增后的人脐带血自然杀伤细胞(hUC-NK)在生物学表型和细胞毒性上的异同。方法:收集健康供者捐赠的脐带血,经淋巴细胞分离液介导的密度梯度离心法富集单个核细胞。... 目的:探讨人脐带血来源的单个核细胞(hUC-MNC),经两种体外培养体系活化扩增后的人脐带血自然杀伤细胞(hUC-NK)在生物学表型和细胞毒性上的异同。方法:收集健康供者捐赠的脐带血,经淋巴细胞分离液介导的密度梯度离心法富集单个核细胞。基于本课题组近期建立的3因子体系(定义为“3IL”),比较Miltenyi和XVIVO 15两种培养基体外培养14 d后NK细胞(分别定义为M-NK和X-NK)的细胞表型、细胞亚群、细胞活力及细胞毒功能的相似性和差异性。结果:经上述2种体系诱导14 d后,总CD3^(-)CD56^(+)NK细胞比例由4.25%±0.04%(d 0)提高至71%±0.18%(M-NK)和75.2%±1.1%(X-NK)。与X-NK组相比,M-NK组CD3^(+)CD4^(+)T和CD3^(+)CD56^(+)NKT细胞比例显著降低。X-NK组中表达CD16、NKG2D、NKp44、CD25活化型标志物的细胞比例更高,但M-NK组总NK细胞扩增数量为X-NK组的2倍。两组在NK细胞增殖及细胞周期上无统计学差异,M-NK组的Annexin V^(+)凋亡细胞比例低于X-NK组。NK细胞与K562细胞体外共培养杀伤实验显示,在相同效靶比(E∶T)下,M-NK组CD107a^(+)NK细胞的比例更高,杀伤效率亦略优于X-NK组(P<0.05)。结论:两种体外扩增活化体系均可诱导产生高活化水平的hUC-NK,但二者在生物学表型和肿瘤杀伤毒性等多个方面存在差异。 展开更多
关键词 NK细胞 活化扩增 细胞亚群 细胞活力 细胞毒性
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脐带间充质干细胞增强伊马替尼诱导慢性粒细胞白血病细胞的凋亡 被引量:3
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作者 刘莹 宋宝全 +5 位作者 魏艺萌 范会芳 余怡 董树旭 韩忠朝 马凤霞 《中国组织工程研究》 CAS 北大核心 2017年第25期4034-4039,共6页
背景:伊马替尼对慢性粒细胞白血病疗效显著,但仍有部分患者对其敏感性较低。人间充质干细胞影响多种血液系统肿瘤的凋亡,但人脐带间充质干细胞是否影响伊马替尼诱导的慢性粒细胞白血病细胞的凋亡的相关研究未见报道。目的:研究人脐带间... 背景:伊马替尼对慢性粒细胞白血病疗效显著,但仍有部分患者对其敏感性较低。人间充质干细胞影响多种血液系统肿瘤的凋亡,但人脐带间充质干细胞是否影响伊马替尼诱导的慢性粒细胞白血病细胞的凋亡的相关研究未见报道。目的:研究人脐带间充质干细胞与伊马替尼联合应用对慢性粒细胞白血病细胞K562细胞凋亡的影响并探讨作用机制。方法:K562细胞与人脐带间充质干细胞和/或伊马替尼共培养。(1)分离培养人脐带间充质干细胞并鉴定;(2)应用流式细胞仪Annexin V-FITC/PI双标法检测细胞凋亡率;(3)Western blot检测凋亡信号通路相关蛋白Bax,Bcl-2,caspase-9,caspase-3,PARP在K562细胞的表达;(4)应用泛caspase抑制剂Z-VAD-FMK抑制caspase信号通路活性,检测caspase凋亡信号通路在人脐带间充质干细胞联合伊马替尼诱导K562细胞凋亡中的作用。结果与结论:(1)体外细胞凋亡实验证实:当人脐带间充质干细胞联合伊马替尼时,K562细胞凋亡率显著增加;(2)Western blot实验证明,当人脐带间充质干细胞联合伊马替尼时,促凋亡蛋白Bax表达升高,抑凋亡蛋白Bcl-2表达降低,caspase凋亡信号通路相关蛋白剪切体形式cleaved-caspase3、cleaved-caspase9及cleaved-PARP均表达升高;(3)泛caspase抑制剂Z-VAD-FMK显著抑制人脐带间充质干细胞和伊马替尼联合应用实验组中K562细胞的凋亡;(4)结果表明,人脐带间充质干细胞联合伊马替尼可促进慢性粒细胞白血病细胞的凋亡,其作用机制是通过激活caspase凋亡信号通路实现的。 展开更多
关键词 干细胞 移植 慢性粒细胞白血病 间充质干细胞 PH染色体 caspase凋亡信号通路 Bax BCL K562细胞 酪氨酸激酶抑制剂
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Effect of intracranial transplantation of CD34^+ cells derived from human umbilical cord blood in rats with cerebral ischemia 被引量:2
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作者 LIU Hai-ying ZhanG Qing-jun +1 位作者 LI Hong-jun han zhong-chao 《Chinese Medical Journal》 SCIE CAS CSCD 2006年第20期1744-1748,共5页
As a source of transplantable stem cells, the CD34^+ subpopulation in human umbilical cord blood (HUCB) has been used extensively to treat some hematopoietic system diseases. However, whether CD34^+ cells hold th... As a source of transplantable stem cells, the CD34^+ subpopulation in human umbilical cord blood (HUCB) has been used extensively to treat some hematopoietic system diseases. However, whether CD34^+ cells hold the therapeutic potential to cerebral ischemia is unknown. The purpose of this study was to observe the recovery of neural function after transplantation of CD34^+ cells derived from HUCB into ischemic cerebral tissue in rats. 展开更多
关键词 fetal blood CD34^+ cell brain ischemia TRANSPLANTATION
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Effect of proline rich domain of an RNA-binding protein Sam68 in cell growth process, death and B cell signal transduction 被引量:1
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作者 LI Qing-hua FAN Tian-xue +2 位作者 PANG Tian-xiang YUAN Wen-su han zhong-chao 《Chinese Medical Journal》 SCIE CAS CSCD 2006年第18期1536-1542,共7页
Background Sam68 plays an important role as a multiple functional RNA binding nuclear protein in cell cycle progress, RNA usage, signal transduction, and tyrosine phosphorylation by Src during mitosis. However, its pr... Background Sam68 plays an important role as a multiple functional RNA binding nuclear protein in cell cycle progress, RNA usage, signal transduction, and tyrosine phosphorylation by Src during mitosis. However, its precise impact on these essential cellular functions remains unclear. The purpose of this study is to further elucidate Sam68 functions in RNA metabolism, signal transduction regulation of cell growth and cell proliferation in DT40 cell line. Methods By using gene targeting method, we isolated a mutation form of Sam68 in DT40 cells and described its effect on cell growth process and signal transduction. Southern, Northern, and Western blot, phosphorylation and flow-cytometric analyses were performed to investigate the Sam68 functions. Results A slower growth rate (2.1 hours growth elongation) and longer S phase (1.7 hours elongation) was observed in the Sam68 mutant cells. Serum depletion resulted in increased amounts of dead cells, and expansion of S phase in mutant cells. Upon B cell cross-linking, the maximal level of tyrosine phosphorylation on BLNK was observed to be significantly lower in mutant cells.Conclusions The proline rich domain of Sam68 is involved in cell growth control by modulating the function of mRNAs in S phase or earlier and the functions as an adaptor molecule in B cell signal transduction pathways. 展开更多
关键词 Sam68 RNA binding protein signal transduction cell death tyrosine phosphorylation
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