肝癌干细胞调控机制的研究进展

肿瘤干细胞学说认为,肝癌的复发和转移主要与肝癌干细胞密切相关.以手术为主的传统治疗肝癌的方法只是杀死了大量快速增殖的肿瘤细胞,并未清除起决定性作用的肝癌干细胞,故术后极易复发转移.调控肝癌干细胞的信号通路及因子较多,如Wnt/β-catenin通路、TGF-β通路、Notch通路、Hedgehog通路、乙型肝炎病毒等.表观遗传学在肝癌干细胞调控机制中的作用亦很重要.深入研究肝癌干细胞的调控机制,可为防止肝癌的复发和转移提供新的治疗依据.     

索拉非尼对肝癌术后早期肝再生的影响

目的研究索拉非尼对肝癌患者术后早期肝再生、肝功能及凝血功能的影响,以及癌旁肝组织的血管内皮生长因子受体2（VEGFR-2）、血管内皮生长因子受体3（VEGFR-3）、血小板衍生的生长因子受体β（PDGFR-β）和原癌基因丝苏氨酸蛋白激酶1（C-Raf-1）的表达水平与肝再生的关系。方法 30例经病理确诊为肝细胞癌的患者,手术切除后接受不同剂量索拉非尼治疗。应用CT测量术后肝脏体积变化,比较不同剂量的索拉非尼对术后1、3个月时肝再生率的影响。分析不同剂量下,服药前及服药后1个月、3个月时患者肝功能及凝血功能的变化。通过免疫组化检测癌旁肝组织中VEGFR-2、VEGFR-3、PDGFR-β和C-Raf-1的表达,分析表达水平与术后1、3个月时肝再生率的相关性。结果索拉非尼服药剂量400 mg/d组的患者术后1个月、3个月时肝再生率平均增加量高于800 mg/d组的患者（P〈0.05）。两组患者服药后3个月时血清AST、ALT明显比服药前升高,血清凝血酶原时间（PT）明显延长（P〈0.05）。术后1、3个月时肝再生率与VEGFR-2、PDGFR-β表达呈负相关（P〈0.05）,VEGFR-2表达对肝再生率的影响最大。结论索拉非尼对肝癌术后早期肝再生有抑制作用,剂量越大抑制效果越明显。VEGFR-2是索拉非尼抑制早期肝再生的主要影响因素。     

肝癌干细胞的研究进展

肿瘤起源于干细胞的假说正在各种人类肿瘤中得到证实.肿瘤不仅是一种基因病,而且是一种干细胞病,基因突变作用于干细胞,干细胞突变成为肿瘤干细胞,这是肿瘤发生、再生、转移和复发的关键.为证实肝癌干细胞的存在,有两个问题必须提出:(1)肝细胞型肝癌是否来源于肝干细胞?(2)肝细胞型肝癌中是否具有干细胞特征的细胞?最新研究表明:肝细胞型肝癌可能是由肝干细胞未分化或分化不全引起的.对于肝癌细胞的研究我们知道肝细胞型肝癌中的细胞具有干细胞特性,如永生性、可移植性以及对治疗手段的抵抗性.但是,至今为止,确切的肝癌干细胞的标记物没有找到,而且没有分离出肝癌干细胞.     

索拉非尼抑制肝癌细胞增殖中自噬的作用及其机制

目的：研究分子靶向药物索拉非尼在体外对人肝癌细胞株HepG_2增殖抑制过程中自噬的表达及作用,并探讨其可能机制。方法：以吖啶橙染色荧光显微镜对自噬进行定性观察;cell counting kit-8检测活性氧（reactiveoxygen species,ROS）抑制前后HepG_2细胞成活率的变化;RT-PCR检测自噬基因Beclin-1表达的变化。Western印迹检测自噬相关蛋白Beclin-1的变化：荧光分光光度计检测胞内二氯荧光素DCF的荧光强度。结果：索拉非尼对肝癌细胞HepG_2具有显著的抑制作用;索拉非尼可诱导肝癌细胞HepG_2产生自噬及ROS,自噬在基因及蛋白水平表达均增加;抑制ROS的产生可减少索拉非尼诱导的肝癌细胞HepG_2自噬的表达量,自噬的抑制增强了索拉非尼对肝癌细胞的抑制作用。结论：ROS参与索拉非尼诱导肝癌细胞HepG_2的自噬表达,索拉非尼在抑制肝癌细胞自噬过程中自噬可能起到保护作用,抑制自噬可能为提高进展期肝癌病人索拉非尼分子靶向治疗敏感性提供新的思路。     

索拉非尼治疗肝癌肝移植术后肿瘤复发的临床疗效观察

探讨索拉非尼治疗原发性肝癌(肝癌)肝移植术后肿瘤复发晚期患者的治疗效果及不良反应.方法 回顾性分析2010年3月至2012年5月郑州人民医院肝脏外科中心收治的9例肝癌肝移植术后肿瘤复发晚期患者的临床资料.确诊肝癌复发后仍然继续原免疫抑制方案,抗肿瘤治疗予索拉非尼口服每次400 mg,每日2次,直至病情进展、发生严重不良反应或死亡中止用药.9例患者每月至少随访1次,全面评价病情,同时检测血清甲胎蛋白(AFP)水平、排斥反应及药物不良反应等情况.结果 9例患者中部分缓解2例,疾病稳定4例,疾病进展3例.用药后2例患者AFP进行性上升,4例患者AFP水平稳定(升高≤10%),3例AFP水平明显下降(下降≥50%).中位肿瘤进展时间为4.2个月,总生存期为6.9个月.不良反应的严重程度主要为1或2级,包括皮疹、腹泻、手足皮肤反应、恶心、呕吐、脱发、食欲减退、乏力、高血压等.经对症处理后患者均能耐受,无1例因药物不良反应而终止治疗.9例患者服用索拉非尼后均未发生排斥反应.结论 索拉非尼作为一种新型多靶向性抗肿瘤药物,对肝癌肝移植术后肿瘤复发的晚期患者有一定的疗效.     

大肠癌干细胞研究进展

随着肿瘤下细胞研究的深入,近年来学者提出了大肠癌起源的干细胞学说,并借助干细胞分离技术,将其成功分离.本篇旨在回顾基于肿瘤干细胞概念提出的大肠癌起源假说和大肠癌干细胞在大肠癌进展的作用,大肠癌干细胞分离技术及其在体内和体外的功能特征,与大肠癌干细胞相关的信号通路及大肠癌耐药机制研究.     

肝细胞肝癌经肝动脉化疗栓塞联合索拉非尼降期后二期切除的初步报告

目的探讨不可一期切除肝细胞肝癌(hepatocellular carcinoma,HCC)经导管肝动脉化疗栓塞(transcatheter hepatic arterial chemoembolization,TACE)联合索拉非尼(Sorafenib)降期治疗后,二期再行根治性切除的可行性。方法回顾性分析2010年3月至2015年1月在南方医科大学南方医院肝胆外科经TACE及口服分子靶向药物索拉非尼治疗、成功降期后再行二期切除的21例HCC病人的临床资料。该组病人平均年龄45.5岁(20~67岁),肝切除手术后持续服用索拉非尼。结果该组病人经TACE联合口服索拉非尼成功降期,降期治疗所需时间平均为52.3 d。降期后实施左、右半肝切除分别为5例和3例,扩大左半肝切除1例,扩大右半肝切除1例,肝脏区段切除11例。术中平均出血量为356.3 ml(150~1 200 ml),平均手术时间为243.3 min(145~365 min)。经过12~62个月随访(中位随访时间为30.1个月),1、2、3年的无瘤存活率分别为76.2%、52.4%、43.6%,复发后病人再次接受TACE、放疗、射频消融等综合治疗者5例,接受再次手术切除的病人3例,1、2、3年总生存率分别为85.7%、71.4%、57.1%。结论 TACE联合口服索拉非尼全身治疗,可使部分初期不可切除肝癌成功降期,降期后接受外科根治性切除手术,初步效果令人鼓舞。     

树突状细胞瘤苗联合吉西他滨对肝癌干细胞的杀伤效应

目的:探讨负载CD133+肝癌细胞抗原的树突状细胞(DC)联合吉西他滨(GEM)在体外对肝癌干细胞的杀伤效应。方法:取对数生长期的人肝癌细胞系Huh-7以CD133作为分子标志进行流式分选,得到肝癌干细胞。将人外周血单个核细胞(PBMC)在体外诱导分化为树突状细胞(DC)。DC负载Huh-7细胞和CD133+细胞抗原后与T细胞共育得到特异性细胞毒性T细胞,将CD133+细胞作为靶细胞进行细胞毒性试验。实验组按处理因素分为:GEM组,CD133+-CTL组,GEM+CD133+-CTL组,Huh7-CTL组和GEM+Huh7-CTL组。CCK-8法检测杀伤率,然后比较各组间差异。结果:对CD133+细胞的杀伤效应以GEM+CD133+-CTL组最强,与其他各组比较差异均有统计学意义(P0.05)。单独就DC瘤苗杀伤率,CD133+-CTL组高于Huh7-CTL组(P0.05)。结论:CD133+肝癌干细胞裂解产物致敏的DC瘤苗联合化疗药物可以有效杀伤肝癌干细胞,进而可能降低肝癌术后和肝癌肝移植后的转移和复发率。     

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目的　明确影响肝癌肝移植术后长期生存率的危险因素 ,探讨符合中国特点的肝癌肝移植适应证。方法　对 1993～ 2 0 0 3年 59例接受单纯肝移植治疗的原发性肝癌病人的移植术前基本资料和肝癌大小、数目、分布以及血管侵犯等属性采用单因素方法进行回顾性分析。结果　对肝癌肝移植术后长期生存有统计学意义的因素包括 :肝功能Child -Pugh分级、AFP水平、肿瘤大小、肿瘤数、门静脉分支侵犯等 (P <0 0 5)以及上腹部手术史 (P <0 0 1)。对肝癌肝移植术后无瘤生存有影响的因素包括上腹部手术史、肿瘤数 (P <0 0 5) ,肿瘤大小和门脉分支侵犯等 (P <0 0 1)。结论　肝功能Child -PughC分级、上腹部手术史和肿瘤侵犯门静脉分支是降低肝癌肝移植术后长期生存率的危险因素 ;而上腹部手术史、肿瘤 >5cm ,数量 >3个和门脉分支侵犯是缩短肝移植术后无瘤生存时间的危险因素。合并肝硬化 ,肿瘤直径≤ 5cm ,数目≤ 3个 ,无侵犯门静脉分支是肝癌肝移植的最佳适应证 ;门静脉分支侵犯应视为肝移植的禁忌证     

索拉菲尼对肝细胞癌凋亡和自噬相关蛋白表达的影响及耐药分析

目的:分析索拉菲尼对肝细胞癌凋亡及自噬相关蛋白表达的影响,及肝细胞癌对其耐药的机制.方法:常规培养人肝癌SMMC-7721细胞,以1.25、2.5、5、10、20μmol/L索拉菲尼处理细胞,观察其对细胞增殖的影响.观察索拉菲尼和自噬抑制剂3-甲基腺嘌呤(3-MA)对细胞凋亡和自噬,及其相关蛋白表达的影响.结果:随着索...     

微小RNA-142-3p靶向CD133对肝癌细胞增殖、侵袭和干细胞特性的影响

目的:观察微小RNA(miR)-142-3p靶向CD133对肝癌细胞增殖、侵袭和干细胞特性的影响。方法:选取郑州大学第二附属医院2017年6月至2019年12月手术切除的肝癌组织和对应的癌旁组织作为研究对象。采用荧光定量聚合酶链反应(PCR)分析癌旁组织和肿瘤组织miR-142-3p表达水平。采用慢病毒在MHCC97H...     

Current status of liver transplantation for hepatocellular cancer

The incidence of hepatocellular cancer is increasing in the United States and is one of the most common cancers worldwide. Traditionally, the gold standard treatment for hepatocellular cancer has been surgical resection, but most patients were not suitable candidates due to advanced disease. Other treatments include locally ablative techniques (cryosurgery, radiofrequency ablation and various injection therapies), chemotherapeutic options and rarely, radiation therapies. In the 1980s, liver transplant emerged as the treatment of choice for end-stage liver disease and also became an option for patients with hepatocellular cancer. When comparing liver transplant with resection in retrospective studies, liver transplant patients had better survival and reduced recurrence. However, not all patients with hepatocellular cancer will be candidates for liver transplant. Size, stage, and histological grade of tumor all affect prognosis after transplant. Use of chemotherapeutic treatments and locally ablative techniques may be beneficial prior to liver transplant, but larger controlled studies are needed. Liver transplant is the most effective treatment for hepatocellular cancer in the subgroup of smaller tumors, but ultimately we are limited by the number of available donors. Future goals in this area include increasing the donor pool and determining optimal management to allow patients to wait for an appropriate donor.     

脂肪间质干细胞抑制肝星状细胞增殖活化及肝脏纤维化

目的 探讨脂肪间质干细胞(adipose tissue-derived mesenchymal stem cells,ADSCs)对活化态肝星状细胞(hepatic stellate cells,HSCs)增殖、活化的影响及其对活体肝硬化模型的治疗作用.方法 分别分离纯化培养HSCs及ADSCs,通过半透膜(transwell insert)建立HSCs与ADSCs的双层培养体系,大鼠正常肝细胞系((buffalo rat liver cells,BRLs)培养作为对照.通过CCK-8比色法检测ADSCs对HSCs增殖的影响;Western blot检测HSCs的α-肌动蛋白(α-SMA)表达;建立鼠肝硬化模型,经门静脉输注ADSCs或BRLs,检测肝组织肝硬化;通过检测培养液中细胞因子的含量探讨可能的作用机制.结果 成功分离获得原代HSCs与ADSCs,活化的HSCs与ADSCs共培养72 h,与对照组相比,ADSCs明显抑制HSCs的活化(各组灰度值分别为1.4±0.2,152±14,258±18,F=283.348,P＜0.05)与增殖(各组吸光度A值分别为2.172±0.107,1.424±0.013,1.209±0.117,F=90.605,P ＜0.05),活体移植显示ADSCs对肝硬化具有明显抑制作用(分别F=77.234、65.164、58.309,均P ＜0.05).培养液细胞因子检测显示ADSCs比BRLs分泌更多的肝细胞生长因子(hepatocyte growth factor,HGF)(F=0.005,P＜0.05),分泌较少的转化生长因子(transforming growth factor-β,TGF-β1)(F=1.767,P＜0.01)及神经生长因子(nerve growth factor,NGF)(F=2.301,P＜0.05). 结论 ADSCs具有分泌细胞因子抑制HSCs活化增殖的潜能,对活体肝硬化进程具有一定的抑制作用.     

氨肽酶N/CD13在肝癌中的表达及意义

目的 探讨氨肽酶N/CD13在肝癌患者中的表达情况,探讨其与肝癌患者临床资料及预后的相关性.方法 应用免疫组化SP法,对40例肝癌患者肝癌组织、10例肝癌患者癌旁组织及10例肝血管瘤患者正常肝组织行单克隆抗体CD13检测.结果 40例(100％)肝癌组织均可见不同程度的CD13阳性表达,肝癌癌旁组织中3例(30％)出现CD13弱阳性表达,10例肝血管瘤患者标本未见表达.CD13表达量与患者的性别、年龄、血清AFP值、HbsAg、分化程度、Child分级及TNM分期差异无统计学意义(P＞0.05);而CD13的表达量与患者的血清AFP值、HbsAg、分化程度之间差异有统计学意义(P＜0.05).生存函数图发现CD13与患者生存时间呈反相关,但CD13表达量与患者肿瘤复发差异无统计学意义(P＞0.05).结论 CD13可作为肝癌干细胞的表面标志物,有望成为判断预后的有效指标.     

骨髓间充质干细胞对人肝癌细胞系MHCC97-H增殖能力的影响

目的 观察骨髓间充质干细胞(MSC)对肝癌细胞增殖能力的影响.方法 人肝癌细胞系MHCC97-H培养液中分别加入0、25%和50%MSC的条件培养基(MSC-CM),采用CyQUANT细胞增殖实验检测吸光度A值变化.在MHCC97-H细胞培养液中添加50%MSC-CM,荧光实时定量聚合酶链反应(RT-PCR)检测肿瘤增殖细胞核抗原(PCNA)、Ki-67抗原的变化.16只裸鼠皮下接种MHCC97-H细胞后,实验组经静脉注射MSC,1×106个/次,每周3次,对照组静脉注射PBS;比较肿瘤体积.结果 培养液中加入MSC-CM后A值依次为211.65±54.72、236.24±57.15和283.59±62.16(P<0.05).PCNA和Ki-67的表达水平分别升高1.8和7.0倍.实验组肿瘤体积(1745±455)mm3m大于对照组(972±568)mm3(P<0.05),肿瘤体积平均增加速度(56.0±26.1)mm3/d高于对照组(32.4±14.5)mm3/d(P<0.05).结论 骨髓间充质干细胞可促进肝癌细胞系MHCC97-H的增殖.     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.     

骨髓源性肝干细胞的筛选、扩增和分化

Objective To explore the feasibility of direct separation, selective proliferation and differentiation of BDLSC from hone marrow cells with culture systems containing cholestatic serum in vitro.Methods Whole bone marrow cells of rats cultured in routine medium were replaced with con-ditioning selection media containing eholestatic sera of different concentrations after they attached to the plates.The optimal concentration of cholestatic serum was determined according to the outcome of the selected cultures.Then the selected BDLSC were induced to proliferating culture system and dif-ferentiating culture system.Each passage of the proliferated stem cells was subjected to flow cytome-try for detection of stem cell markers.The morphology and phenotypie markers of BDLSC were char-acterized using immunohistochemistry, RT-PCR and electron microscopy.The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.Results The 50 ml/L eholestatie serum was the optimal concentration for the selection of BDLSC at which BDLSC could sur-vive while the other populations of the bone marrow cells could not.The purified BDLSC could prolif-erate for 6 passages and maintained stable markers in our proliferating system.When the culture sys-tem changed to differentiating system, hepatocyte-like colony-forming units (HCFU) were formed.HCFU expressed markers of embryonic hepatoeytes (AFP, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and eytoehrome P450-2b1), and hepatocyte nuclear factors (HNF-1α and HNF-3β).They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.Conclusion BDLSC can be selected directly from whole bone marrow cells and pure BDLSC can also proliferate for 6 passages.The differentiated cells have hepatocyte-like phenotype and function.BDLSC will he a new way to provide a readily avail-able alternate source of cells for clinical hepatocyte therapy.