CD133:A cancer stem cells marker, is used in colorectal cancers

Colorectal cancer is one of the most common malignant tumors worldwide. A model of cancer development involving cancer stem cells has been put forward because it provides a possible explanation of tumor hierarchy. Cancer stem cells are characterized by their proliferation, tumorigenesis, differentiation, and selfrenewal capacities, and chemoradiotherapy resistance. Due to the role of cancer stem cells in tumor initiation and treatment failure, studies of cancer stem cell markers, such as CD133, have been of great interest. CD133, a five-transmembrane glycoprotein, is widely used as a marker to identify and isolate colorectal cancer stem cells. This marker has been investigated to better understand the characteristics and functions of cancer stem cells. Moreover, it can also be used to predict tumor progression, patient survival, chemoradiotherapy resistance and other clinical parameters. In this review, we discuss the use of CD133 in the identification of colorectal cancer stem cell, which is currently controversial. Although the function of CD133 is as yet unclear, we have discussed several possible functions and associated mechanisms that may partially explain the role of CD133 in colorectal cancers. In addition, we focus on the prognostic value of CD133 in colorectal cancers. Finally, we predict that CD133 may be used as a possible target for colorectal cancer treatment.

Autologous CD34^+ and CD133^+ stem cells transplantation in patients with end stage liver disease

AIM:To assess the utility of an autologous CD34 + and CD133 + stem cells infusion as a possible therapeutic modality in patients with end-stage liver diseases.METHODS:One hundred and forty patients with endstage liver diseases were randomized into two groups.Group 1,comprising 90 patients,received granulocyte colony stimulating factor for five days followed by autologous CD34 + and CD133 + stem cell infusion in the portal vein.Group 2,comprising 50 patients,received regular liver treatment only and served as a control group.RESULTS:Near normalization of liver enzymes and improvement in synthetic function were observed in 54.5% of the group 1 patients;13.6% of the patients showed stable states in the infused group.None of the patients in the control group showed improvement.No adverse effects were noted.CONCLUSION:Our data showed that a CD34 + and CD133 + stem cells infusion can be used as supportive treatment for end-stage liver disease with satisfactory tolerability.

Upregulated CD133 expression in tumorigenesis of colon cancer cells

AIM: To analyze the upregulated CD133 expression in tumorigenesis of primary colon cancer cells. METHODS: Upregulated CD133 expression in tumorigenesis of colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) was analyzed by flow cytometry. Human colon cancer tissue samples were stained with anti-human CD133. SW620 cells were sorted according to the CD133 expression level measured by fluorescence-activated cell sorting. Spheroids of colorectal cancer cells were cultured with the hanging drop. Expression of CD133 and Lgr5 in spheroids of colorectal cancer cells and monolayer culture was detected by RT-qPCR. Spheroids of colorectal cancer cells were analyzed using anti-human CD133 with immunohistochemical staining. RESULTS: CD133 antigen was expressed in colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) as well as in primary and metastatic human colon cancer tissues. However, the CD133 was differently expressed in these cell lines and tissues. The expression levels of CD133 and Lgr5 were significantly higher in spheroids of parental, CD133hi and CD133-cells than in their monolayer culture at the mRNA level (P < 0.05). Immunohistochemical staining of spheroids of CD133-cells showed that CD133 was highly expressed in colorectal cancer cell lines. CONCLUSION: Upregulated CD133 expression plays a role in tumorigenesis colorectal cancer cells, which may promote the expression of other critical genes that can drive tumorigenesis.

CD133^+ gallbladder carcinoma cells exhibit self-renewal ability and tumorigenicity

AIM： To identify cancer stern cells （CSCs） in human gallbladder carcinomas （GBCs）. METHODS： Primary GBC cells were cultured under serum-free conditions to produce floating spheres. The stem-cell properties of the sphere-forming cells, including self-renewal, differentiation potential, chemoresistance and tumorigenicity, were determined in vitro or in vivo. Cell surface expression of CD133 was investigated in primary tumors and in spheroid cells using flow cytometry. The sphere-colony-formation ability and tumorigenicity of CD133＋ cells were assayed.floating spheroids were generated from primary GBC cells, and these sphere-forming cells could generate new progeny spheroids in serum-free media. Spheroid cells were differentiated under serum-containing conditions with downregulation of the stem cell markers Oct-4, Nanog, and nestin （P 〈 0.05）. The differentiated cells showed lower spheroid-colony-formation ability than the original spheroid cells （P 〈 0.05）. Spheroid ceils were more resistant to chemotherapeutic reagents than the congenetic adherent cells （P 〈 0.05）. Flow cytometry showed enriched CD133＋ population in sphereforming cells （P 〈 0.05）. CD133＋ cells possessed high colony-formation ability than the CD133 population （P 〈 0.01）. CD133＋ cells injected into nude mice revealed higher tumorigenicity than their antigen-negative counterparts （P 〈 0.05）. CONCLUSION： CD133 may be a cell surface marker for CSCs in GBC.

Inhibitive effect of IL-24 gene on CD133^+ laryngeal cancer cells

Objective:To explore the inhihitive and apoptosis inductive effect of IL-24 genes on CD133^+laryngeal cancer cells in Hep-2 line.Methods:Human peripheral blood monocytes were isolated.The total RNA was extracted by using Trizol method and reverse transcripted into cDNA using RT-PCR method.Primers P1 and P2 was designed for the amplification of human IL-24 genes.After confirmation of agarose gel electrophoresis tests,TA was cloned into pMD19-T simple vector.Nhe Ⅰ and Xho Ⅰ double digesting human IL-24 and pIRES2-ZsGreen1 and eukaryotic expression vector were used to establish the pIRES2-ZsGreen1-hIL-24 vector,and detected by enzyme digestion and gene sequencing methods.Flow cytometry(FCM) was used to isolate CD133^+ cells from Hep-2 cells.CD133^+ cells were transfected with pIRES2-ZsGreen1-hIL-24 through liposome 2000.After detection,MTT and FCM were used to observe the effect of IL-24 gene on CD133^+ laryngeal cancer Hep-2 cells.Results:Lipotin mediated transfection of recombinant pIRES2-ZsGreen1-hIL-24 plasmid into CD133^+ Hep-2 could expressed IL-24 gene in cells stably.MTT results showed that IL-24 transfected group was significantly suppressed compared to empty vector group and control group(P<0.05);FCM results showed that the apoptosis rate of experimental group increased significantly compared to empty vector group and control group(P<0.05).Conclusions:IL-24 gene expressions can inhibit proliferation of CD133^+laryngeal cells in Hep-2 line and promote their apoptosis.

Angioarchitecture and CD133^+ tumor stem cell distribution in intracranial hemangiopericytoma A comparative study with meningioma

Angioarchitecture plays an important role in the malignant development of intracranial hemangiopericytoma. It remains poorly understood whether high frequency of hemorrhage during clinical surgery for intracranial hemangiopericytoma is associated with angioarchitecture. The present study utilized hematoxylin-eosin staining, and immunohistochemical staining with epithelial membrane antigen, vimentin, CD34, von Willebrand factor （vWF） and CD133 to observe characteristics of angioarchitecture. In addition, silver stains were used to demonstrate changes in reticular fibers in the wall of vessel channels in intracranial hemangiopericytoma and meningioma. Five patterns of angioarchitecture were identified in intracranial hemangiopericytoma, namely tumor cell islands, vasculogenic mimicry, mosaic blood vessels, sprouting angiogenesis, and intussusceptive angiogenesis. Several CD133＋ tumor cells were found to form tumor cell islands. A connection between vWF ^＋ and vWF channels was detected in the pattern of intussusceptive angiogenesis, and some vimentin^＋ tumor cells were embedded in the periodic acid-Schiff positive channel wall. Incomplete threads of reticular fibers formed the walls of larger pseudo-vascular channels and some tumor clumps or scattered tumor cells were detected ＂floating＂ in them. The angioarchitecture, specific markers and reticular fibers of intracranial hemangiopericytoma were significantly different from meningioma. Angioarchitecture provides a functional vascular network for vascular evolution in intracranial hemangiopericytoma and contributes to significant intra-operative bleeding.

WJSC 6^(th) Anniversary Special Issues(2):Mesenchymal stem cells Intestinal stem cells and celiac disease

Stem cells(SCs)are the key to tissue genesis and regeneration.Given their central role in homeostasis,dysfunctions of the SC compartment play a pivotal role in the development of cancers,degenerative disorders,chronic inflammatory pathologies and organ failure.The gastrointestinal tract is constantly exposed to harsh mechanical and chemical conditions and most of the epithelial cells are replaced every 3 to 5 d.According to the so-called Unitarian hypothesis,this renewal is driven by a common intestinal stem cell(ISC)residing within the crypt base at the origin of the crypt-to-villus hierarchical migratory pattern.Celiac disease(CD)can be defined as a chronic immune-mediated disease that is triggered and maintained by dietary proteins(gluten)in genetically predisposed individuals.Many advances have been achieved over the last years in understanding of the pathogenic interactions among genetic,immunological and environmental factors in CD,with a particular emphasis on intestinal barrier and gut microbiota.Conversely,little is known about ISC modulation and deregulation in active celiac disease and upon a gluten-free diet.Nonetheless,bone marrow-derived SC transplantation has become an option for celiac patients with complicated or refractory disease.This manuscript summarizes the"state of the art"regarding CD and ISCs,their niche and potential role in the development and treatment of the disease.

Cisplatin selects for CD133+cells in lung cancer cells

Objective Platinum-based chemotherapy is the first-line treatment for non-small cell lung cancer,but the chemoresistance of tumor cells continues to be a considerable challenge in the management of NSCLCs,leading to recurrence of most patients.CD133(prominin-1)is a five-transmembrane glycoprotein,and recent evidence suggests that CD133+cells are the cause of drug resistance and tumor recurrence.In this study,the correlation between cisplatin and CD133+cells was investigated systematically.Methods Four lung cancer cell lines,including A549,H460,801D and H1299,were treated with different concentrations of cisplatin.Cell viability was determined by MTT assay.Sphere-forming assay was performed to detect the capability of sphere-forming.CD133+cells was detected by BD FACScaliber flow cytometer.Results The results showed that cisplatin could increase the number of CD133+cells in both time-and dose-dependent manner.The enrichment would weaken but the proportion of CD133+cells was still higher than the basic level as incubation time extended after cisplatin was withdrawn.Compared with adherent culture,the proportion of CD133+cells was higher when the cells were maintained suspension culture.The proportion of CD133+cells significantly increased when cisplatin was provided in suspension culture.Conclusion These results revealed that cisplatin induces the enrichment of CD133+cells and CD133 is a new therapeutic target.Our data partially explained drug resistance to second-line chemotherapy in cisplatin-treated patients with NSCLCs.

The Cross-talk between ROS and p38MAPKα in the Ex Vivo Expanded Human Umbilical Cord Blood CD133^+ Cells

This study investigated the correlation between and compared the effects of reactive oxygen species（ROS） and p38 mitogen-activated protein kinase α（p38MAPKα） in the ex vivo expanded umbilical cord blood（hUCB） CD133＋ cells.hUCB CD133＋ cells were cultured in the hematopoietic stem cells（HSCs） culture medium with N-acetylcysteine（NAC,an anti-oxidant）,p38MAPKα-specific inhibitor（SB203580） or their combination.The levels of ROS and expression of phosphorylated p38MAPKα（p-p38） in CD133＋ cells were flow cytometrically detected.The efficacy of ex vivo expansion was evaluated by the density of CD133＋ cell sub-group colony-forming cells（CFC） and cobblestone area-forming cells（CAFC） assay.Our results showed decreased ROS levels in NAC,SB203580,and their combination treatment groups were almost 37%,48%,and 85%,respectively.Furthermore,SB203580 abrogated the activation of p38MAPKα more obviously than NAC.Moreover,the CD133＋ cells in SB203580 treatment group had a 21.93±1.36-fold increase,and 14.50±1.19-fold increase in NAC treatment group,but only 10.13±0.57-fold increase in control group.In addition,SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did.These findings suggested that,in expanded CD133＋ cells,ROS activates p38MAPKα,which,in turn,induces ROS production,and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion.

Influence of CD133^+ expression on patients' survival and resistance of CD133^+ cells to anti-tumor reagents in gastric cancer

Objective:To investigate the influence of CD133+expression on patients'survival and resistance of CD133+cells to anti-tumor agents in gastric cancer(GC).Methods:Influence of CD133 expression on prognosis was analyzed employing samples from patients with GC.GC cell lines were utilized to separate CD133+and CD133-subpopulations by immunomagnetic separation and to analyze the biological features of two subpopulations in vitro and in vivo,especially in resistant to anti-tumor reagents and its apoptotic mechanism.Results:The lower CD133+group showed a significantly better survival compared with the higher CD133+group.The highest content of CD133+subpopulations for KATO-III cells had stronger proliferative ability than CD133-subpopulations.A single CD133+cell was capable of generating new cell colony and the tumorigenicity rate in nude mice was100%for CD133+clonal spheres or for CD133+cells,but 0%for CD133-cells.Furthermore,the higher expression levels of Oct-4,Sox-2,Musashi-1 and ABCG2 in CD133+clonal spheres were identified compared with CD133+cells or CD133-cells.Under the treatment of anti-tumor reagents,CD133+cells had lower suppression rates compared with CD133-cells while lower level of Bcl-2 and higher level of Bax were found in CD133+cells compared with CD133-cells.Conclusions:The patients with lower CD133+expression had a better survival.Enriched CD133+cells in clonal sphere shared the ability to be self-renewable,proliferative,tumorigenic and resistant to anti-tumor agents as probably regulated by Bcl-2 and Bax.

Identification and analysis of a minority fraction in a U87 cell line Do side-population cells represent bona fide stem cell-like cancer cells?

BACKGROUND： Overwhelming evidence suggests that tumor bulks are comprised of differentiated tumor cells and cancer stem cells （CSCs）. The stem cell-like side-population （SP） cells account for a minor fraction of the total tumor cells, yet are apparently the cells capable of tumor initiation, growth, maintenance, and recurrence. OBJECTIVE： To identify potential stem cell-like cancer cells in a U87 human brain glioma cell line on the basis of dye efflux, clone formation, and multi-drug resistance capacity. DESIGN, TIME AND SETTING： The cellular and molecular biology experiment was performed at the Laboratory of Shanghai Institute of Hematology and Laboratory of Shanghai Institute of Endocrinology in Ruijin Hospital; in vivo contrast observational animal trial was performed at Experimental Animal Center, School of Medicine, Shanghai Jiao Tong University from June 2007 to May 2008. MATERIALS： The U87 cell line was provided by the Shanghai Institute of Cancer Research, Chinese Academy of Science; DMEM/F12 （1 ： 1） and fetal bovine serum were purchased from Gibco Invitrogen, USA; human recombinant basic fibroblast growth factors were purchased from BD Bioscience, USA; Hoechst 33342, Verapamil, and methyl thiazolyl tetrazolium were purchased from Sigma, USA; phycoerythrin-labeled anti-human-CD133 was purchased from Milteny Biotec, Germany; SYBR PrimeScriptTM RT-PCR kit was purchased from TaKaRa Biotechnology, Dalian, China. METHODS： Monolayer cultured cells were harvested by 0.25% Trypsin-EDTA and suspended at a 1 ×10^6/mL dilution in PBS containing 2% FBS, and were stained with Hoechst 33342 dye, either alone or in combination with Verapamil. Following fluorescence-activated cell sorting, SP and non-SP subsets were cultivated with serum-containing （DMEM plus 10% fetal bovine serum） or serum-free culture medium [DMEM/F12 （1： 1） ＋ 1× B27 supplement ＋ 10 ng/mL basic fibroblast growth factors ＋ 1× L-glutamine] to determine growth characteristics in vitro. Finally, single free U87 cells and subsets （SP or non-SP cells） were subcutaneously injected into the backs of 5-week-old nude mice for in vivo tumorigenicity. MAIN OUTCOME MEASURES： Cell morphology and clonogenicity were observed under inverted microscope; SP phenotype and fluorescent antibody labeling were analyzed by MoFIoTM flow cytometry; ABC transporter mRNA expression was evaluated by semi-quantitative real-time RT-PCR; efflux capacity for anti-neoplastic drugs from the U87 cell line and subsets was measured with the MTT assay, then detected by enzyme-linked immunosorbent assay at a wavelength of 490 nm; in vivo tumorigenicity in immunodeficient nude mice was evaluated by diameter size. RESULTS： During in vitro passages, human U87 cells maintained a stable SP fraction profile and exhibited the ability to form neurosphere-like clones. SP cell proliferation decreased compared with non-treated U87 cells. CD133 expression was reduced in the SP and non-SP cells. Freshly sorted SP fractions expressed higher levels of ABC drug transporter genes, and exhibited increased potential for cytotoxic drug resistance. The in vivo malignancy of U87 cells was largely dependent on non-SP cells in nude mice, and tumors that formed from the non-SP fraction developed faster and larger compared with tumors from the SP fraction. CONCLUSION： The SP cell component was a key factor that influenced mRNA expression and cytotoxic drug resistance. In particular, cancer stem cells or tumor-initiating cells were not exclusively enriched in the SP subset of the U87 cell line, and non-SP cells were even more tumorigenic.

Proliferation Characteristics of CD133+ Cell Population in Colorectal Cancer

In this study,CD133+ subpopulations were isolated from 41 primary colorectal cancer tissues,the proliferation and cell cycle distribution of the cells were examined without in vitro expansion,and then compared to those of cell lines.The detection of CD133 in colorectal cancer tissues,isolation of CD133+ and CD133-epithelial subpopulations,Ki-67/DNA multiparameter assay and cell volume analysis were flow cytometrically conducted.The results showed that Ki-67 expression was correlated with CD133 level in primary cancer tissues,while cell cycle G 2 /M phase distribution or clinicopathological characteristics was not.In addition,the CD133+ cells showed larger cell volume and higher Ki-67 expression as compared with CD133-cells.But there was no statistically significant difference in G 2 /M phase distribution between the two subpopulations.Our results demonstrated that the CD133+ subpopulation in colorectal cancer tissue contained more actively cycling and proliferating cells,which was not correlated to clinicopathological factors but might contribute to tumor progression and poor clinical outcome.

Influence on Biological Behavior of Colon Cancer Stem Cells after RNA Interfering CD133

OBJECTIVE To observe the effects expression and activation on biological cancer stem cells. of blocking CD133 gene characteristic of the colon METHODS CD133＋ colon cancer stem cells （CCSCs） were separated from EpCAMhigh CD44＋ CCSCs through fluorescenceactivated cell sorting （FACS）. The proliferation, the capability of spherical cell formation, neoplasia, and the expression of ABCG2 mRNA of CD133＋ CCSCs were observed after the CD133＋ CCSCs were infected with LV-CD133shRNA. CD133 negative cells were isolated from EpCAMhigh CD44＋ CCSCs with FACS, and the CD133 proteins in CD133- cells were detected with Western blot. RESULTS CD133＋ CCSCs were isolated from EpCAMhigh CD44＋ CCSCs using FACS, and they accounted for 89.2% in the stem cells. In the experimental group, after the CD133＋ CCSCs were knocked down by LV-CD133shRNA RNAi, the growth pattern of the cells in the stem cell culture changed into adherent growth from suspended growth, and couldn＇t generate spherical cells. Results of MTT assay showed that the CD133＋ CCSCs infected with LV-CD133shRNA grew slowly, compared to the cells in the control groups. There was a decrease in the cloning efficiency. The infected cells were transplanted into the BALB/c nude mice. During the observation, no neoplasia was found in the CD133＋ cells infected with LV-CD133shRNA. The level of ABCG2 mRNA expression was lowered greatly （P 〈 0.01）. CD133- cells were obtained from the EpCAMhigh CD44＋ CCSCs using FACS, in which the expression of CD133 protein was positive. CONCLUSION CD133 retains the biological characteristics of the colon cancer stem cells.

Isolation and characterization of cancer stem-like cells from MHCC97H Cell Lines

Objective：To identify and isolate CD133 positive cancer stem-like cells （CD133^＋ cells） from the highly invasive human hepatocellular carcinoma cell Iine（MHCC97H）, and examine their potential for clonogenicity and tumorigenicity. Methods： CD133^＋ and CD133^- cells were isolated from MHCC97H cell line by magnetic bead cell sorting（MACS）, and the potentials of CD133^＋ cells for colony formation and tumorigenicity were evaluated by soft agar cloning and tumor formation following nude mice inoculation. Results：CD133^＋ cells represent a minority（0.5-2.0%） of the tumor cell population with a greater colony-forming efficiency and greater tumor production ability. The colony-forming efficiency of CD133^＋ cells in soft agar was significantly higher than CD133^- cells（36.8 ± 1.4 vs 12.9 ± 0.8, P 〈 0.05）. After 6 weeks, 3/5 mice inoculated with 1 × 10^3 CD133^＋ cells, 4/5 with 1 × 10^4 CD133^＋ cells and 5/5 with 1× 10^5 CD133^＋ cells developed detectable tumors at the injection site, while only one tumor was found in mice treated with same numbers of CD133 cells. Conclusion： CD133 may be a hallmark of liver cancer stem cells （CSC） in human hepatocellular carcinoma（HCC）, because the CD133^＋ cells identified and isolated with anti-CD133 labeled magnetic beads from MHCC97H cell line exhibit high potentials for clonogenicity and tumorigenicity. These CD 133^＋ cells might contribute to hepatocarcinogenesis, as well as the growth and recurrence of human HCC, and therefore may be a useful target for anti-cancer therapy.

低氧训练对大鼠心肌组织血管内皮祖细胞标记AC133表达的影响

目的:探讨低氧训练对大鼠心肌组织血管内皮祖细胞标记AC133表达的影响。方法:健康雄性SD大鼠16只,随机分为2组:对照组、高住高练组。高住高练组采用10周递增负荷跑台运动训练,每周训练6天,运动量由第1周的速度为15m/min、持续时间为25min递增至第10周速度为28m/min、持续时间为50min,每周二、四、六在相当于海拔1500m的低氧环境中训练,并且在低氧环境中居住,低氧程度由第1周相当于海拔1800m递增至第10周相当于海拔3600m。采用免疫组织化学方法检测CD34,定位心肌组织微血管;采用免疫组织化学及免疫电镜方法,检测微血管上AC133蛋白表达。结果:CD34可较好地标记心肌组织微血管;对照组大鼠心肌组织微血管内皮上有微弱的AC133蛋白表达,高住高练组大鼠心肌组织微血管内皮上有较多的AC133蛋白表达。结论:大鼠心肌内皮祖细胞归巢参与血管新生可能是低氧训练促进心肌血管新生的机制之一。

AC133抗原在急性白血病细胞中的表达

为了解AC133抗原在急性白血病细胞中的表达情况及其临床意义 ,采用常规三色直接免疫荧光标记法 ,使用流式细胞仪测定AC133抗原及其它白血病相关抗原在 61例急性白血病患者细胞中的表达。检测结果显示 ,61例中有 2 7例 (44 .3% )AC133表达阳性 ,其中ALL 13例 (13 2 9) ,AML M0 M16例 (6 8) ,AML M2 4例 (4 16) ,AML M4 M54例 (4 8)。 2 7例AC133+病例中2 5例 (92 .6% )同时表达CD34,13例 (48.1% )同时表达CD117。本研究结果提示 ,AC133抗原与CD34和CD117的表达有相关性。AC133抗原可表达于各型白血病中 ,但在AML M0

翼状胬肉组织中CD_(34)、AC133、STRO-1、C-KIT的表达

目的评估源自骨髓的干细胞与翼状胬肉发病的相关性,探寻翼状胬肉的发病机制。方法选择翼状胬肉头部达到瞳孔缘的病例12例(12眼),其中男6例,女6例;年龄51～66岁;原发性翼状胬肉8例,复发性翼状胬肉4例,均长于鼻侧。对照组材料来自同一眼球、距翼状胬肉边缘约10mm的正常球结膜组织。采用EnVision免疫组织化学法检测骨髓干细胞标志物来源的CD34、AC133、STRO-1、C-KIT的表达。结果光镜下可见翼状胬肉组织中有大量带有骨髓干细胞或祖细胞阳性标志物来源的细胞,翼状胬肉组织头部阳性表达更强。4种阳性细胞的分布及细胞形态类似。C-KIT阳性表达细胞聚集于翼状胬肉上皮基底组织和基质组织。上皮基底部阳性细胞呈圆形或椭圆形,而在基质组织中呈纺锤形且主要分布于血管周围。CD34阳性表达细胞主要分布于翼状胬肉组织上皮基底层,在血管内皮也有阳性表达,类似于C-KIT,但比C-KIT上皮层组织阳性细胞少。细胞形态在上皮基底部呈圆形或椭圆形。AC133阳性表达细胞主要分布于上皮层,基质和血管内皮中也有阳性表达,形态类似CD34和C-KIT。STRO-1阳性表达细胞的分布不同,主要见于基质层,阳性细胞形态呈纺锤形,而上皮层中未见表达。各种因子的表达在原发性和复发性翼状胬肉间无明显差别。对照组切片中标志物的表达均呈阴性。结论带有源自骨髓干细胞阳性标志物的细胞在翼状胬肉的发病和术后复发过程中可能起重要作用。

脐血来源AC133^+细胞在不同诱导条件下神经分化标志物的表达(英文)

背景:近年来,在啮齿类动物中发现,造血干细胞具有向神经分化的塑型性,而在人类,这种造血的塑型性仍然具有争议。目的:检测人脐血来源的AC133+造血干细胞是否具有向神经分化的潜能。设计、时间及地点:本实验为成组对照设计实验,于2005-08/2007-12在天津血液学研究所和清华大学玉泉医院神经中心实验室完成。材料:人脐带取自健康足月新生儿。胎脑滋养层细胞来源于22周流产胎儿脑组织。方法:应用免疫磁珠分选人脐血AC133+细胞,流式细胞仪检测其纯度为99%以上。同时以机械分离及酶消化法制备胎脑滋养层细胞。选用4种培养条件对脐血来源AC133+造血干细胞进行诱导分化:①生长培养液DMEM/F12加上皮生长因子和碱性成纤维生长因子。②DMEM/F12加上皮生长因子和碱性成纤维生长因子并联合使用脑源性生长因子,其间用全反式维甲酸处理3d。③非细胞接触的共培养体系,先将制备的胎脑滋养层细胞生长在培养板内插件的半透膜上,与培养板内的脐血来源的AC133+细胞进行共培养。④细胞直接接触共培养,将预先用BrdU标记的脐血来源的AC133+细胞直接种在胎脑滋养层细胞上进行共培养。主要观察指标:1,2,4周收集诱导分化的脐血细胞,以RT-PCT检测是否有巢蛋白、骨形态生成蛋白2及神经细胞黏附分子的表达,免疫细胞化学方法检测细胞分型特异性抗原。结果:部分脐血来源的AC133+细胞,在体外存在生长必需的因子上皮生长因子和碱性成纤维生长因子时,能表达一些与神经细胞分化有关的基因,如巢蛋白、骨形态生成蛋白,条件不同时,这些基因出现下调和关闭。在DMEM/F12加上皮生长因子和碱性成纤维生长因子联合使用脑源性生长因子诱导培养液中,上述基因表达在2周时可检测到,同时可检测到神经发育过程中出现的另一分子神经细胞黏附分子,这一分子在造血过程中并不出现,在使用内插件的胎脑滋养层细胞共培养的脐血细胞中也检测到相同结果。在优化的非细胞接触条件中主要表达胶质酸性蛋白,在细胞直接接触的共培养体系中出现神经元分化的标志物Ⅲβ-tubulin蛋白的表达。这种基因表达变化提示,在特定的培养环境下,脐血来源的AC133+细胞内可能出现了基因重排,使造血潜能的干细胞表达神经细胞发育分子。结论:脐血来源的AC133+细胞包含部分具有向神经分化潜能的多能干细胞,在适合条件下,表达神经分化相关抗原。

AC133-2分子的克隆及其逆转录病毒表达载体的构建

目的克隆人AC133-2全长基因,构建PGEZ-Term-AC133—2逆转录病毒表达载体。方法采用分段克隆的方法,通过聚合链式反应从胎肝文库中克隆AC133-2,并构建AC133—2基因的逆转录病毒表达载体。结果成功克隆AC133—2全长基因并构建PGEZ-Term-AC133-2逆转录病毒表达载体。结论AC133-2全长基因克隆及构建逆转录病毒表达载体的成功,为构建AC133-2转基因细胞和制备抗人AC133-2单克隆抗体和研究AC133-2的生物学功能奠定了物质基础。