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1.
本研究主要目的是明确M-CSF诱导骨髓间充质干细胞分化为肝样细胞的分子机制,为临床中的肝移植和治疗肝病提供新思路。对取自于本院骨科治疗的患者的股骨骨髓间充质干细胞进行提取、分离、传代培养及鉴定。流式细胞仪检测BMSCs的表面表型。为了诱导BMSCs的肝分化,本研究将BMSCs加入到培养基中。骨髓间充质干细胞诱导21 d后,BMSCs表达了肝细胞特异性标志物a-蛋白(AFP)和细胞角蛋白18(CK18),通过免疫荧光染色证实了分化与为分化的BMSCs表达的差异性。分化的BMSCs还显示了肝细胞的体外功能特征,包括白蛋白产生、尿素分泌和糖原储存。本研究结果表明,BMSCs在M-CSF诱导下可分化为功能性肝细胞样细胞,可作为肝病治疗的细胞来源。 相似文献
2.
Bone marrow-derived mesenchymal stem cells (BMSCs) are of particular interest in the field of tissue engineering because of
their potential to differentiate into osteoblasts, chondrocytes, and neuronal cells. In order to promote the differentiation
of BMSCs into specific cell types, appropriate scaffold biomaterials and bioactive molecules that can support the differentiation
of BMSCs into specific cell types are needed. We hypothesized that β-mercaptoethanol (BME), which has been reported to induce
the differentiation of BMSCs into neural-like cells, promotes BMSCs to differentiate into neural-like cells when BME is added
to polymeric scaffolds containing the BMSCs. We fabricated biocompatible film shaped scaffolds composed of poly(lacti-co-glycolic)
acid (PLGA) and various concentrations of BME to confirm that BME-promoted differentiation of BMSCs is concentration-dependent.
Cell proliferation increased as the BME concentration in the films increased at the early stage, and the proliferation rate
remained similar on the PLGA films for 3 weeks following the BMSC seeding. The expression of neuronal markers in differentiated
BMSCs was assessed by RT-PCR. At 2- and 3-week time-points, mRNA expression of neurofilament and neuron specific enolase was
significantly increased in PLGA/BME films containing 400 μM BME compared to PLGA films. Thus, we have identified BMSC-seeded
PLGA/BME films with 200 μM and 400 μM BME as potentially useful candidates for neural tissue engineering applications by promoting
BMSC proliferation and differentiation towards neural-like cells. 相似文献
3.
Endothelial cells and smooth muscle cells (SMCs) are important aspects of vascularization in vaginal reconstruction. Research has confirmed that mesenchymal stem cells could differentiate into endothelial-like cells and SMCs. But the methods were more complicated and the mechanism was unknown. In the current study, we induced the bone mesenchymal stem cells (BMSCs) to differentiate into endothelial-like cells and SMCs in vitro by differentiation medium and investigated the effect of Wnt/β-catenin signaling on the differentiation process of BMSCs. Results showed that the hypoxic environment combined with VEGF and bFGF could induce increased expression of endothelial-like cells markers VEGFR1, VEGFR2, and vWF. The SMCs derived from BMSCs induced by TGF-β1 and PDGF-AB significantly expressed SMC markers SMMHC11 and α-SMA. The data also showed that activation of Wnt/β-catenin signaling could promote the differentiation of BMSCs into endothelial-like cells and SMCs. Thus, we established endothelial-like cells and SMCs in vitro by more simple methods, presented the important role of hypoxic environment on the differentiation of BMSCs into endothelial-like cells, and confirmed that the Wnt/β-catenin signaling pathway has a positive impact on the differentiation of BMSCs into endothelial-like cells and SMCs. This is important for vascular reconstruction. 相似文献
4.
A novel in vitro retinal differentiation model by co-culturing adult human bone marrow stem cells with retinal pigmented epithelium cells 总被引:4,自引:0,他引:4
Chiou SH Kao CL Peng CH Chen SJ Tarng YW Ku HH Chen YC Shyr YM Liu RS Hsu CJ Yang DM Hsu WM Kuo CD Lee CH 《Biochemical and biophysical research communications》2005,326(3):578-585
Human retinal pigment epithelium (HRPE) cells are important in maintaining the normal physiology within the neurosensory retina and photoreceptors. Recently, transplantation of HRPE has become a possible therapeutic approach for retinal degeneration. By negative immunoselection (CD45 and glycophorin A), in this study, we have isolated and cultivated adult human bone marrow stem cells (BMSCs) with multilineage differentiation potential. After a 2- to 4-week culture under chondrogenic, osteogenic, adipogenic, and hepatogenic induction medium, these BMSCs were found to differentiate into cartilage, bone, adipocyte, and hepatocyte-like cells, respectively. We also showed that these BMSCs could differentiate into neural precursor cells (nestin-positive) and mature neurons (MAP-2 and Tuj1-positive) following treatment of neural selection and induction medium for 1 month. Furthermore, the plasticity of BMSCs was confirmed by initiating their differentiation into retinal cells and photoreceptor lineages by co-culturing with HRPE cells. The latter system provides an ex vivo expansion model of culturing photoreceptors for the treatment of retinal degeneration diseases. 相似文献
5.
Fereshteh Azedi Somaieh Kazemnejad Amir Hassan Zarnani Masoud Soleimani Amir Shojaei Shaghayegh Arasteh 《Molecular biology reports》2017,44(1):169-182
In order to characterize the potency of menstrual blood stem cells (MenSCs) for future cell therapy of neurological disorders instead of bone marrow stem cells (BMSCs) as a well-known and conventional source of adult stem cells, we examined the in vitro differentiation potential of these stem cells into neural-like cells. The differentiation potential of MenSCs to neural cells in comparison with BMSCs was assessed under two step neural differentiation including conversion to neurosphere-like cells and final differentiation. The expression levels of Nestin, Microtubule-associated protein 2, gamma-aminobutyric acid type B receptor subunit 1 and 2, and Tubulin, beta 3 class III mRNA and/or protein were up-regulated during development of MenSCs into neurosphere-like cells (NSCs) and neural-like cells. The up-regulation level of these markers in differentiated neural-like cells from MenSCs was comparable with differentiated cells from BMSCs. Moreover, both differentiated MenSCs and BMSCs expressed high levels of potassium, calcium and sodium channel genes developing functional channels with electrophysiological recording. For the first time, we demonstrated that MenSCs are a unique cell population with differentiation ability into neural-like cells comparable to BMSCs. In addition, we have introduced an approach to generate NSCs from MenSCs and BMSCs and their further differentiation into neural-like cells in vitro. Our results hold a promise to future stem cell therapy of neurological disorders using NSCs derived from menstrual blood, an accessible source in every woman. 相似文献
6.
摘要 目的:研究大鼠BMSCs(骨髓间充质干细胞)原代培养与纯度鉴定的方法。方法:无菌环境中,从SD大鼠股骨与胫骨端采集骨髓,先行酶消化,利用全骨髓细胞悬液贴壁法对提取BMSCs实施传代培养,选取生长良好的第3代细胞进行鉴定;对BMSCs实施成脂与成骨诱导分化,同时经由油红O(ORO)与茜素红(ARS)染色法对诱导分化效果加以鉴定;借助流式细胞术(FCM)对CD34、CD44与CD90这3类BMSCs表面标志物的表达展开分析。结果:BMSCs是长梭状贴壁细胞,生长状态为纤维细胞样漩涡状;在第3代BMSCs传代期间,其第1-3 d发展至生长潜伏期,呈较慢速的生长;第3-5 d发展至对数生长期,呈高速生长;待至第7 d长速增殖最大,速度停止上升进入平缓期;BMSCs成骨、成脂诱导结束后,对其诱导分化鉴定发现:细胞出现明显形态学变化,通过ORO对脂肪染色,细胞显示橘红色;待成骨诱导培养结束,通过ARS对钙盐染色,显示红色,且出现矿化结节沉积,说明BMSCs具有良好的成骨、成脂分化能力;FCM测定发现:CD34表达呈阴性(1.09 %),CD90(96.8 %)与CD44(92.4 %)皆呈阳性,与BMSCs表型相符。结论:经由全骨髓黏附培养技术有效分离BMSCs,且完成培养。 相似文献
7.
骨髓间充质干细胞又称为骨髓源性间充质干细胞,是指存在于骨髓基质细胞系统中的一类干细胞,具有高度稳定的体外扩增能力和多向分化潜能等特点。骨髓间充质干细胞因其取材方便,易于分离和培养,以及在适当条件下可诱导分化为皮肤、骨骼、内脏、血液、神经等多种组织细胞的独特优势,目前被广泛应用于药物开发、免疫调节、组织修复、器官重建等多个研究领域。近年来,骨髓间充质干细胞作为种子细胞在组织工程领域有着非常诱人的潜在应用前景。本文就骨髓间充质干细胞在组织工程学研究中应用的最新进展作一综述。 相似文献
8.
Mesenchymal stem cells (MSCs) can differentiate into neural cells to treat nervous system diseases. Magnetic resonance is an ideal means for cell tracking through labeling cells with superparamagnetic iron oxide (SPIO). However, no studies have described the neural differentiation ability of SPIO-labeled MSCs, which is the foundation for cell therapy and cell tracking in vivo. Our results showed that bone marrow-derived mesenchymal stem cells (BM-MSCs) labeled in vitro with SPIO can be induced into neural-like cells without affecting the viability and labeling efficiency. The cellular uptake of SPIO was maintained after labeled BM-MSCs differentiated into neural-like cells, which were the basis for transplanted cells that can be dynamically and non-invasively tracked in vivo by MRI. Moreover, the SPIO-labeled induced neural-like cells showed neural cell morphology and expressed related markers such as NSE, MAP-2. Furthermore, whole-cell patch clamp recording demonstrated that these neural-like cells exhibited electrophysiological properties of neurons. More importantly, there was no significant difference in the cellular viability and [Ca2+]i between the induced labeled and unlabeled neural-like cells. In this study, we show for the first time that SPIO-labeled MSCs retained their differentiation capacity and could differentiate into neural-like cells with high cell viability and a good cellular state in vitro. 相似文献
9.
Expression of hepatocyte-like phenotypes in bone marrow stromal cells after HGF induction 总被引:33,自引:0,他引:33
Wang PP Wang JH Yan ZP Hu MY Lau GK Fan ST Luk JM 《Biochemical and biophysical research communications》2004,320(3):712-716
Bone marrow comprises heterogeneous cell populations, of which certain progenitors have demonstrated the ability to differentiate into multiple mesenchymal cell lineages. This study demonstrates the bone marrow stromal cells (BMSCs) with intrinsic plasticity to differentiate into hepatocyte-like phenotypes under in vitro induction of hepatocyte growth factor (HGF). BMSCs isolated from rat femurs and tibias were cultured and passaged 3-4 times in the presence of HGF. Cells were harvested on days 0, 10, and 20 and subjected to examination of any hepatocyte characteristics by flow cytometry, RT-PCR, Western blot, and immunocytochemistry. Expression of albumin and alpha-fetoprotein at both mRNA and protein levels was detectable on day 10. By contrast, c-Met mRNA was significantly decreased in BMSC in the course of HGF induction. Here BMSC was shown to differentiate into hepatocyte-like phenotypes given the HGF induction, as an alternative source for adult stem cell transplantation in liver repair. 相似文献
10.
Jian-Dong Yang Cheng-Huang Jing-Cheng Wang Xin-Min Feng Yi-Nan Li Hai-Xiang Xiao 《Cytotechnology》2014,66(6):1007-1019
The bone marrow represents the most common source from which to isolate mesenchymal stem cells (MSCs). They can be obtained directly from patients and successfully induced to form various differentiated cell types. In addition, cell-based transplantation therapies have been proven to be promising strategies for curing disease of the nerve system. Therefore, it was particularly important to establish an easy and feasible method for the isolation, purification, and differentiation of bone marrow stromal cells (BMSCs). The aim of this study was to isolate and characterize putative bone marrow derived MSCs from Sprague–Dawley (SD) rats. Furthermore, differentiation effects were compared between the GDNF-induction group and the BDNF-induction group. Of these, BMSCs were isolated from the SD rats in a traditional manner, and identified based on plastic adherence, morphology, and surface phenotype assays. After induction with GDNF and BDNF, viability of BMSCs was detected by MTT assay and neuronal differentiation of BMSCs was confirmed by using immunofluorescence and Western blotting. Besides, the number of BMSCs that obviously exhibited neuronal morphology was counted and the results were compared between the GDNF-induction group and BDNF-induction groups. Our results indicate that direct adherence was a simple and convenient method for isolation and cultivation of BMSCs. Furthermore, BMSCs can be induced in vitro to differentiate into neuronal cells by using GDNF, which could achieve a more persistent and stable inducing effect than when using BDNF. 相似文献
11.
12.
Wanyao Xia Yu-Qing Jin James D. Kretlow Wei Liu Wenlong Ding Hengyun Sun Guangdong Zhou Wenjie Zhang Yilin Cao 《Biotechnology letters》2009,31(5):639-646
TGF-β1 plays a necessary and important role in the induction of chondrogenic differentiation of bone marrow stromal cells
(BMSCs). In this study, porcine BMSCs were infected with a replication-deficient adenovirus expression vector carrying the
hTGF-β1 gene. The transduced BMSCs were cultured as pelleted micromasses in vitro for 21 days, seeded onto disk-shaped PGA
scaffolds for 3 days and subsequently implanted into the subcutaneous tissue of mice. BMSCs transduced with AdhTGF-β1 expressed
and secreted more hTGF-β1 protein in vitro than those of the control group. Histological and immunohistological examination
of the pellets revealed robust chondrogenic differentiation. Tissues made from cells transduced with AdhTGF-β1 exhibited neocartilage
formation after 3 weeks in vivo. The neocartilage occupied 42 ± 5% of the total tissue volume which was significantly greater
than that of the control group. Furthermore, there was extensive staining for sulfated proteoglycans and type II collagen
in the AdhTGF-β1 group compared to controls, and quantification of GAG content showed significantly greater amounts of GAG
in experimental groups. The results demonstrate that transfer of hTGF-β1 into BMSCs via adenoviral transduction can induce
chondrogenic differentiation in vitro and enhance chondrogenesis in vivo. 相似文献
13.
Magda M. El-Mahdi Wafaa A. Mansour Olfat Hammam Noha A. Mehana Taghreed M. Hussein 《The Korean journal of parasitology》2014,52(2):151-162
The technique of stem cells or hepatocytes transplantation has recently improved in order to bridge the time before whole-organ liver transplantation. In the present study, unfractionated bone marrow stem cells (BMSCs) were harvested from the tibial and femoral marrow compartments of male mice, which were cultured in Dulbecco''s modified Eagle''s medium (DMEM) with and without hepatocyte growth factor (HGF), and then transplanted into Schistosoma mansoni-infected female mice on their 8th week post-infection. Mice were sacrificed monthly until the third month of bone marrow transplantation, serum was collected, and albumin concentration, ALT, AST, and alkaline phosphatase (ALP) activities were assayed. On the other hand, immunohistopathological and immunohistochemical changes of granuloma size and number, collagen content, and cells expressing OV-6 were detected for identification of liver fibrosis. BMSCs were shown to differentiate into hepatocyte-like cells. Serum ALT, AST, and ALP were markedly reduced in the group of mice treated with BMSCs than in the untreated control group. Also, granuloma showed a marked decrease in size and number as compared to the BMSCs untreated group. Collagen content showed marked decrease after the third month of treatment with BMSCs. On the other hand, the expression of OV-6 increased detecting the presence of newly formed hepatocytes after BMSCs treatment. BMSCs with or without HGF infusion significantly enhanced hepatic regeneration in S. mansoni-induced fibrotic liver model and have pathologic and immunohistopathologic therapeutic effects. Also, this new therapeutic trend could generate new hepatocytes to improve the overall liver functions. 相似文献
14.
HSV-1感染大鼠骨髓间充质干细胞及形成潜伏感染的初步研究 总被引:2,自引:0,他引:2
在体外培养大鼠骨髓间充质干细胞(BMSCs),观察单纯疱疹病毒1型感染骨髓间充质干细胞情况.分离并鉴定BMSCs;HSV-1感染BMSCs,观察细胞病变(CPE);建立BMSCs的HSV-1潜伏感染模型.提取总DNA,PCR法扩增BMSCs内的HSV-1特异性片段,检测HSV-1感染BMSCs及潜伏感染.结果显示骨髓间充质干细胞经14d诱导后,碱性磷酸酶含量增高、形成钙结节,表现出成骨细胞特性.HSV-1感染BMSCs,出现典型的CPE,PCR法证实BMSCs内存在HSV-1的特异性片段.HSV-1潜伏感染的BMSCs,未出现明显的CPE,细胞传至7代,仍可测到HSV-1的基因片段,表明BMSCs有可能形成HSV-1的潜伏感染.大鼠骨髓间充质干细胞在体外可以向成骨细胞方向分化,可作为组织工程学的种子细胞.HSV-1可以在体外感染骨髓间充质干细胞并有形成潜伏感染的趋势. 相似文献
15.
Bone marrow mesenchymal stem cells (BMSCs) have been shown to differentiate into cells of a neural lineage. However, no studies have examined whether gender influences the differentiation potential of BMSCs. Here, we explore the possible differences in BMSC’s neurogenic potential in vitro between female rhesus monkey BMSCs (F-rhBMSCs) and male rhBMSCs (M-rhBMSCs). We first isolated and cultured rhBMSCs from female and male donors (n = 6, 2 years old), identified their sex origin by karyotype assay, and assessed their expression of nestin and CD34 at passage 1 and 10. Then, nestin-positive F- and M-rhBMSCs at P10 were differentiated into neural-like cells. After induction, the neurogenic potential of these cells was assessed by morphological observation and protein expression analysis of neural markers, including class III β-tubulin, neurofilament light polypeptide, tau, and gamma-aminobutyric acid (GABA) neurotransmitter. Furthermore, GABA content was assayed using high-pressure liquid chromatography. The results showed that F-rhBMSCs produced significantly more nestin-positive cells compared with M-rhBMSCs at P10 and that nestin-positive F-rhBMSCs acquired higher neurogenic potential accompanied by increased synthesis and excretion of GABA compared with nestin-positive M-rhBMSCs under conditions of differentiation. These results indicated that gender may play an important role in the neurogenic potential of BMSCs, and a further understanding of the cellular biology underlying these differences may contribute to the development of new therapeutic strategies for neurological repair and regeneration. 相似文献
16.
It has been reported that rat bone marrow stromal cells (BMSCs) are differentiated into neuronal cells by administration of
2-mercaptoethanol [Woodbury et al (2000) J Neurosci Res 61:364–370]. In this study, we examined the effects of various sulfhydryl
(SH) compounds on the differentiation of BMSCs obtained from rat femurs. Neuronal differentiation was detected morphologically
and immunocytochemically. It was found that the cells treated with reduced glutathione (GSH) apparently differentiated into
neurons, showing extensive processes, and expressing neuron-specific enolase and microtubule-associated protein 2. Glutathione
monoethyl ester (GEE), which increased the cellular GSH content, showed no effect on the expression of neuronal markers. It
is concluded that the neural differentiation of BMSCs occurs by the administration of GSH. It was suggested that extracellular
and not intracellular GSH have effects on the induction of the neuronal differentiation of BMSCs. 相似文献
17.
骨髓基质干细胞诱导分化为神经元过程中miR-124和miR-128的表达 总被引:1,自引:0,他引:1
目的探讨骨髓基质干细胞诱导分化为神经元过程中miR-124和miR-128的表达变化及作用。方法采用全骨髓培养法体外分离培养获得骨髓基质干细胞,取传代培养至第3代的骨髓基质干细胞,在神经干细胞培养液及细胞因子等条件下诱导其分化为神经元,倒置显微镜下观察其形态变化,应用ABI公司的TaqManMicroRNAAssaysreal-timePCR技术,检测miR-124和miR-128在诱导分化过程中的表达。结果 miR-124分化后神经元的表达是未分化BMSCs的0.051倍(P0.05);miR-128分化后神经元的表达是未分化BMSCs的0.070倍(P0.05)。结论 miR-124和miR-128在骨髓基质干细胞诱导分化为神经元过程中可能起重要作用。 相似文献
18.
Dong-Xiang Qian Hong-Tian Zhang Xu Ma Xiao-Dan Jiang Ru-Xiang Xu 《Neurochemical research》2010,35(4):572-579
The aim of this study was to compare the neural differentiation potential and the expression of neurotrophic factors (NTFs) in differentiated adipose-derived stem cells (ADSCs) using three established induction protocols, serum free (Protocol 1), chemical reagents (Protocol 2), and spontaneous (Protocol 3) protocols. Protocol 1 produced the highest percentage of mature neural-like cells (MAP2ab+). Protocol 2 showed the highest percentage of immature neural-like cells (β-tubulin III+), but the neural-like state was transient and reversible. Protocol 3 caused ADSCs to differentiate spontaneously into immature neural-like cells, but not into mature neural cell types. The neural-like cells produced by Protocol 1 lived the longest in culture with little cell death, but Protocol 2 and 3 led to the significant cell death. Therefore, Protocol 1 is the most efficient among these protocols. Additionally, soon after differentiation, the mRNA levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in dADSCs were sharply decreased by Protocol 1 and 2 (acute induction protocol), but not by Protocol 3 (chronic induction protocol). The results indicate that NTFs played an important role in neural differentiation via acute responses to NGF and BDNF, but not chronically during the transdifferentiation process. 相似文献
19.
Zhiqiang Liu Jingda Xu Haiyan Li Yuhuan Zheng Jin He Huan Liu Yuping Zhong Yong Lu Bangxing Hong Mingjun Zhang Pei Lin Juan Du Jian Hou Jianfei Qian Larry W. Kwak Qing Yi Jing Yang 《PloS one》2013,8(12)
Multiple myeloma (MM) cells are responsible for aberrant osteoclast (OC) activation. However, when cocultured monocytes, but not OC precursors, with MM cells, we made a novel observation that MM cells inhibited receptor activator of nuclear factor κB ligand (RANKL)-induced increase of OC differentiation, OC gene expression, signaling pathways and bone resorption activity. Our results showed that MM cells produced multiple inhibitory cytokines of osteoclastogenesis, such as IL-10, which activated STAT3 signaling and induce OC inhibition. However, cocultures of bone marrow stromal cells (BMSCs) reversed MM-induced OC inhibition. We found that MM cells increased production of MCP-1 from BMSCs and BMSC-derived MCP-1 enhanced OC formation. Mechanistic studies showed that IL-10 downregulated RANK expression in monocytes and thus, inhibited RANKL-induced OC formation. In contrast, MCP-1 upregulated RANK expression and thus, enhanced OC formation. Overall, our studies for the first time demonstrated that MM cell have inhibitory effects on osteoclastogenesis by producing inhibitory cytokines. Our results further indicate that activation of osteoclastogenesis in bone marrow requests the crosstalk of MM cells, BMSCs and their produced cytokines. Thus, our studies provide evidences that targeting bone marrow microenvironmental cells and/or cytokines may be a new approach to treating MM bone destruction. 相似文献
20.
Hideki Agata Nobukazu Watanabe Noriyuki Kubo Mika Yamazaki Hideaki Kagami 《Biochemical and biophysical research communications》2009,382(2):353-155
Current standard techniques for bone tissue engineering utilize ex vivo expanded osteogenic cells. However, ex vivo expansion requires serum, which may hinder clinical applications. Here, we report the feasibility and efficacy of bone tissue engineering with human bone marrow stromal cells (BMSCs) expanded in serum-free conditions. Bone marrow was aspirated from 4 healthy donors and adherent cells were cultured in either serum-free medium (STEMPRO® MSC SFM) or conventional serum-containing medium (α-MEM supplemented with 10% serum). Efficacy of expansion was greater in serum-free medium. Phenotypically, serum-free expanded BMSCs were smaller in cell-size and showed expression of CD105++ and CD146dim. After osteogenic induction, serum-free expanded BMSCs showed lower alkaline phosphatase activity. However, they showed higher responsiveness to induction. In vivo bone-forming ability was also confirmed. In conclusion, bone tissue engineering with serum-free expanded BMSCs is feasible and as efficient as that obtained with BMSCs expanded in conventional serum-containing medium. 相似文献