改进型纤维蛋白凝胶种植技术在组织工程骨体外构建中的应用

目的探讨纤维蛋白凝胶种植技术在组织工程骨体外构建中的应用,从而改善目前常规构建技术的缺陷。方法首先采用逆转录病毒pLEGFP-N1对人骨髓间充质干细胞进行荧光蛋白标记,经成骨诱导培养作为种子细胞,然后分别应用纤维蛋白凝胶种植技术(A组)和沉淀种植技术(B组),体外构建细胞-材料复合体,通过倒置荧光显微镜和扫描电镜等观测种子细胞在支架材料内的分布与数量。结果逆转录病毒载体pLEGFP-N1成功标记骨髓间充质干细胞,并应用于工程化组织体外构建的示踪;A组细胞在材料内分布更均匀、数量更多,构建第2天细胞数量为(6.83±0.15)×105/个复合体,而B组细胞为(3.28±0.18)×105/个复合体,两组数据有非常显著统计学意义(P<0.01)。结论纤维蛋白凝胶种植技术简单易行,不仅改善种子细胞在骨支架材料内的分布,而且提高细胞种植效率,为体外高效构建组织工程骨创造了条件。     

细胞共培养技术及其在骨软骨组织工程中的应用

目的：探讨细胞共培养技术及其在骨软骨组织工程中的应用。方法：在CNKI数据库和Pubmed中检索近年来有关细胞共培养技术及其在骨软骨组织工程中应用的相关文献,进行汇总分析。结果：细胞共培养技术诱导干细胞向体细胞分化、维持细胞的功能和活力、对细胞增殖进行调控,有效地扩大了骨软骨损伤修复所需的种子细胞源。结论：细胞共培养技术充分模拟了体内环境,使细胞间能相互沟通信息,相互支撑生长增殖,使其成为组织工程领域一种新兴而很有发展前景的技术。     

脂肪干细胞在骨软骨组织工程中的研究进展

近年来,随着医学相关学科的迅猛发展,组织工程技术已经取得了很大的成就,但种子细胞一直是困扰人们的焦点问题.脂肪干细胞(ADSCs)体外扩增迅速,稳定性好,无免疫排斥反应,具有多向分化潜能,可以向脂肪细胞、成骨细胞、成软骨细胞、内皮细胞、肌细胞和神经细胞等不同胚层来源的细胞分化,并且脂肪组织具有取材方便、对人体创伤小、可大量获得等优点,因而ADSCs有望成为一种理想的组织工程种子细胞.旨在介绍ADSCs在骨软骨组织工程中的研究进展.     

骨髓间充质干细胞及其在骨/软骨缺损修复中的应用进展

间充质干细胞是一类多能干细胞，在胚胎形成过程中分化为骨、软骨、肌腱、肌肉、脂肪和髓基质等多种间充质组织。近年，研究者成功地分离到人和多种动物骨髓间充质干细胞并发现其在体外仍保持干细胞特性，能够诱导分化为成骨细胞、软骨细胞、成肌细胞和脂肪细胞等细胞系。动物实验表明，骨髓间充质干细胞能够修复具有临床意义的软骨缺损。其在肌腱缺损修复中的应用价值与巳初步得到证实。此外，由于具有多能性，间充质干细胞还是很好的基因载体。在创伤修复的基因治疗中有广阔的应用前景。     

组织工程化干细胞联合PRP在骨不连中的临床应用

目的 比较骨不连患者采用组织工程干细胞联合PRP和常规开放固定+有效植骨治疗的临床疗效。方法 将骨不连患者43例分为治疗组22例和对照组21例,治疗组患者采用组织工程化干细胞联合PRP移植治疗,对照组采用开放内固定+有效植骨治疗。比较患者两组手术时间、出血量、住院费用、住院时间、切口愈合等级、有无取自体骨等相关指标。结果 随访9~15个月后,治疗组骨折愈合率为86.36%,对照组骨折愈合率为85.70%,组间比较,差异无统计学意义（P＞0.05）;在手术时间、出血量、住院费用、住院时间方面,对照组均高于治疗组,差异有统计学意义（P＜0.05）;切口愈合方面,对照组有2例患者出现Ⅱ级愈合,治疗组患者均为Ⅰ级愈合;有无取自体骨:治疗组无一例取自体骨,对照组均取自体骨。结论 组织工程化干细胞联合PRP可作为一种有效的治疗骨不连的方法,较常规手术方法更微创,操作更简便,损伤小,节约经济,减少住院时间,未来可作为治疗骨不连的一种有效的方法。     

骨髓间充质干细胞在骨组织工程中的研究进展

近20年来，组织工程取得了迅猛发展，为骨缺损修复提供了新的思路和治疗途径，要构建理想的的组织工程化骨其前提就是要有生物学特性良好的种子细胞和适合的支架材料，随着干细胞研究的不断深入，给组织工程种子细胞的选择带来了新的希望。目前，可应用于骨组织工程的种子细胞中，骨髓间充质干细胞（bone marrow mesenchymal stem cells，MSCs），因其可通过体外贴壁培养分离，扩增迅速，是一类具有分化潜能的细胞，在特定条件下可以分化为多种组织细胞，如成骨细胞、软骨细胞、肌腱、脂肪细胞、成纤维细胞以及神经星状细胞等，且具有极强的自我复制能力，是一种重要组织工程种子细胞。本文就骨髓间充质干细胞作为骨组织工程的种子细胞的国内外研究进展简要做一综述。     

rFN／CDH—BCP组织工程骨的制备和体内融合效应的初步研究

目的制备复合bMSCs的rFN／CDH—BCP组织工程骨,并对其体内融合效应进行初步观测。方法制备复合bM-SCs的rFN／CDH—BCP组织工程骨,利用细胞离心粘附实验、MTT实验和成骨诱导分化等方法观察组织工程骨界面对骨种子细胞的粘附和分化的影响。建立兔腰椎横突间植骨融合模型,以复合bMSCs的rFN／CDH-BCP组织工程骨植于预备的骨槽内,利用X线影像学和组织学技术观察融合部位新骨生成状况及种子细胞分布、转归状况。结果bMSCs复合24h后rFN／CDH—BCP表面和内部有大量细胞粘附,生长良好,成骨诱导10d后,rFN／CDH-BCP表面ALP活性显著升高（P〈0．05）,21d后,钙结节广泛分布。bMSCs复合rFN／CDH-BCP植入兔植骨融合区12周后X线和组织学显示有明显新生骨生成,对照组没有。结论rFN／CDH—BCP组织工程骨在体内外均有良好的生物学效应,对新骨形成和骨成熟具有显著的促进作用和良好修复作用,具有较广阔的临床应用前景。     

骨髓间充质干细胞复合鸡蛋膜构建组织工程化骨的实验研究

目的探讨鸡蛋膜作为组织工程支架材料同大鼠骨髓间充质干细胞（bone marrow mesenchy-cmal stem cells,BMSCs）复合培养构建组织工程化骨的可行性。方法采用梯度离心法分离大鼠骨髓间充质干细胞;将第2代BMSCs同鸡蛋膜支架材料复合,利用含0.1μmol/L地塞米松、10mmol/Lβ-甘油酸钠、50μg/ml维生素C的条件培养液诱导培养2周后,分别将复合材料植入Wistar大鼠右侧背部皮下,左侧植入无细胞复合的鸡蛋膜作为对照。28d后处死大鼠收集标本,利用扫描电镜技术、HE染色、免疫组织化学（Ⅰ型胶原、碱性磷酸酶）和茜素红染色进行相应检测。结果细胞同鸡蛋膜复合培养后,细胞在鸡蛋膜上容易贴附生长,7d后鸡蛋膜表面有大量的细胞生长,14d后细胞连接成片且鸡蛋膜网状孔隙中有大量的细胞生长。复合材料植入大鼠28d后,实验组鸡蛋膜内部有大量细胞存在,植入区可见有组织形成,对照组鸡蛋膜内部无细胞,植入区未见组织形成;实验组BMSCs向成骨细胞分化并分泌矿化基质,表达Ⅰ型胶原,碱性磷酸酶,对照组均为阴性。结论鸡蛋膜具有良好的组织生物相容性,以BMSCs作为种子细胞、鸡蛋膜作为支架材料构建组织工程化骨具备一定的可行性。     

3种组织来源的间充质干细胞体外成骨能力的比较

目的比较成人骨髓间充质干细胞(BMSCs)、人脐带间充质干细胞(UC-MSCs)和人胎盘间充质干细胞(P-MSCs)的成骨能力。方法用含10%胎牛血清的DMEM/Ham's F-12培养液培养3种MSCs,CCK8法检测增殖能力,流式细胞仪鉴定3种细胞。碱性磷酸酶(ALP)和茜素红染色观察细胞经成骨诱导后成骨分化蛋白-ALP的分泌和矿化钙结节的沉积。实时荧光定量PCR(RT-q PCR)法检测MSCs骨再生相关基因的表达。Western blot方法检测MSCs成骨再生相关基因的蛋白表达。结果 MSCs在第3天进入对数增殖期。3种细胞的表面标志物阳性率:CD44、CD90和CD105均高于98%。3种MSCs成骨诱导9 d时,3种MSCs的实验组均表达大量成骨分化蛋白-ALP,成骨诱导18 d时3种MSCs均呈现较好的矿化能力;3种MSCs成骨诱导9 d时,实验组RUNX2和ALP基因显著性高表达(P0.05),成骨诱导18 d时,实验组RUNX2和骨钙素(OCN)亦显著性高表达(P0.05);3种MSCs成骨诱导9 d时,实验组均检测到RUNX2和ALP的蛋白表达;成骨诱导18 d时,实验组细胞亦检测到RUNX2和OCN的蛋白表达。结论 UC-MSCs和P-MSCs具有良好的成骨分化能力,有望作为骨组织工程的种子细胞用于治疗骨缺损。     

同种异体骨髓间充质干细胞在比格犬体内异位构建组织工程骨

BACKGROUND: Allogenic bone marrow mesenchymal stem cells as seed cells for tissue engineering have become the future trend of development. OBJECTIVE: To investigate the osteogenic effects of allogenic bone marrow mesenchymal stem cells and the outcome in vivo. METHODS: Bone marrow mesenchymal stem cells from beagle dogs were marked with chloromethylbenzoyl ammonia fluorescent dye (CM-Dil), and the proliferation of labeled cells was measured using MTT assay in vitro. Autologous or allogenic bone marrow mesenchymal stem cells were inoculated into coral and β-tricalcium phosphate scaffolds for 7 days osteogenic induction and then subcutaneously implanted into the back of beagle dogs. Dogs undergoing blank scaffold implantation served as negative controls. Hematoxylin-eosin staining was used to observe new bone formation at 3 days, 1, 2, 4, 8, 12 weeks after surgery. Bone formation area was statistically analyzed using ipp software. In the CM-Dil group, frozen sections were made to trace the in vivo outcome of bone marrow mesenchymal stem cells under a fluorescence microscope. RESULTS AND CONCLUSION:The osteogenesis speed in the allogenic bone tissue engineering group was faster than that in the autologous bone tissue engineering group at 4-8 weeks after implantation, but no significant difference between the two groups was found beginning at the 12th week. At 4 weeks after implantation, the expression of γ-carboxy glutamic acid protein in the autologous bone tissue engineering group was higher than that in the allogenic bone tissue engineering group, prompting the bone mineralization appeared earlier in the latter group than the former one. ELISA results showed that the expression of alkaline phosphatase and osteocalcin in the autologous bone tissue engineering group was higher than that in the allogenic bone tissue engineering group at 4 weeks after implantation, and then the expression showed no difference at 12 weeks. CM-Dil labeling results showed that the number of allogenic bone marrow mesenchymal stem cells was reduced significantly compared with that of autologous bone marrow mesenchymal stem cells. All these findings indicate that the ectopic osteogenesis of the allogenic tissue-engineered bone in large animals is found within 12 weeks after implantation, but the osteogenesis efficiency at early stage (within 8 weeks) is lower compared with the autologous tissue-engineered bone. This difference may be related to the post-implantation immunoreactions that lead to the reduction in cell number. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

细胞传代对骨髓间充质干细胞向神经干细胞分化的影响

BACKGROUND:It is unclear whether serial cell passage in vitro influences the differentiation of bone marrow mesenchymal stem cells into neural stem cells. OBJECTIVE:To investigate the effect of cell passage on the differentiation of bone marrow mesenchymal stem cells into neural stem cells. METHODS:Rat bone marrow mesenchymal stem cells were isolated and cultured by the whole bone marrow adherence method. Bone marrow mesenchymal stem cells at passages 3, 6, 9, 12 were incubated in serum-free medium. After culture for 7 and 14 days, cell biological characterization was observed and differenitaiton ability into neural stem cells was observed by detecting Nestin expression in cells using flow cytometry. Then, the cells were further induced to differentiate and cell multipotential differentiation capacity was detected by measurement of nerve enolase and glial acidic protein expression. RESULTS AND CONCLUSION:Under induction, bone marrow mesenchymal stem cells at different passages were all differentiated into Nestin-positive neural stem cells. However, there was a significant difference in differentiation proportion of cells at different passages (P < 0.05). Strongest differentiation ability was found in the passage 6 cells, with the Nestin expression up to (93.7±2.3)% at 7 days of induction and (96.2±1.8)% at 14 days of induction. The proportion of differentiated cells at passages 6 and 9 was signfiicantly higher than that at passages 3 and 12. Moreover, adherent cells were positive for nerve enolase and glial acidic protein. All these findings indicate that the differentiation of bone marrow mesenchymal stem cells into neural stem cells is correlated with cell passage. Cells at lower or higher passages are both detrimental to cell differentiation.     

小儿骨髓间质干细胞体外诱导分化为神经元样细胞

BACKGROUND:In the past, the culture and differentiation of bone marrow mesenchymal stem cells in vitro were mostly reported in the adult or animal rather than in children. OBJECTIVE:To explore the ability of bone marrow mesenchymal stem cells from children differentiating into neural stem cells and nerve cells. METHODS:Bone marrow mesenchymal stem cells from children were isolated and cultured, and passage 12 cells were cultured in the pre-induction medium (DMEM culture medium containing 10% fetal bovine serum and 1 mmol/L β-mercapto ethanol) and induction medium (DMEM containing 2% dimethyl sulfoxide and 150 μmol/L butylated hydroxyanisole). Expression of nestin and β-tublin III was detected using immunocytochemistry method at 30 minutes and 7 days after induction, while RT-PCR was used to detect nestin mRNA expression at 0, 5.5, 6 days after induction. RESULTS AND CONCLUSION: After combined induction, the cells shrank from round shape to tapered, polygonal or oval shape, and cell processes extended gradually and became filament-like shape. Interconnected cells formed a network at 6 days after combined induction. The expression of nestin antigen was positive at 30 minutes after induction, while the expression of β-tublin was positive at 7 days. RT-PCR findings showed that positive expression of nestin mRNA was detected at 5.5 hours of induction, and then disappeared at 6 days. These findings show that the combined use of dimethyl sulfoxide and butylated hydroxyanisole can induce bone marrow mesenchymal stem cells from children to differentiate into neural stem cells and nerve cells in vitro.     

腺病毒携带骨形态发生蛋白14基因转染脂肪干细胞修复损伤关节软骨

背景：关节软骨损伤后自我修复能力较弱,主要是由于其缺乏滋养血管并且细胞代谢缓慢等组织特性,目前的治疗方法都不能恢复软骨组织的原有功能,近年来软骨组织工程已引起了越来越多的关注。 目的：观察Ⅰ型胶原海绵支架搭载骨形态发生蛋白14基因转染脂肪干细胞修复兔膝关节软骨损伤的效果。 方法：取兔皮下脂肪组织分离培养脂肪干细胞,用腺病毒真核表达载体Ad-CMV-BMP-14-IRES-hrGFP-1转染脂肪干细胞。Ⅰ型胶原海绵支架搭载转染后的脂肪干细胞,待细胞吸附后对兔膝关节全层软骨缺损进行修复。术后12周取手术关节,从大体方面、组织学方面综合评估缺损修复状况。 结果与结论：骨形态发生蛋白14转染后的脂肪干细胞骨形态发生蛋白14和Ⅱ型胶原蛋白表达及Sox-9基因表达明显高于普通脂肪干细胞。术后12周,支架搭载经骨形态发生蛋白14转染的脂肪干细胞组软骨组织修复良好,平整光滑,光洁度、质地及颜色良好,交界区整合良好。支架搭载脂肪干细胞组软骨组织部分修复,有正常软骨光泽,质地与颜色接近正常,修复组织与正常软骨组织界限明显。单纯支架组几乎崩解塌陷,未见透明样软骨结构形成。结果可见腺病毒携带骨形态发生蛋白14基因转染后脂肪干细胞修复软骨缺损的能力有大幅提升。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

骨髓间充质干细胞调节心脏移植大鼠的免疫功能

BACKGROUND:Heart transplantation is an effective method for treatment of end-stage heart failure, but immune rejection that seriously impact therapeutic effacicy is easy to occur after transplantation. OBJECTIVE:To investigate the regulatory effect of bone marrow mesenchymal stem cells on the immune function of rats undergoiong heart transplantation. METHODS:Twenty Lewis rats were enrolled as donors, and 20 Wistar rats as recipients. Heart transplantation models were established in the Wistar rats. These 20 model rats were randomized into cell transplantation and control group with 10 rats in each group. Forty-eight hours after heart transplantation, rats in the cell transplantation group were given bone marrow mesenchymal stem cell suspension (1 mL, 2×10⁸ cells/L) via the tail vein, while rats in the control group were given normal saline in the same dose. Then, the expression levels of serum interleukin-2, interleukin-10 and percentage of CD4⁺, CD8⁺, CD4⁺/CD8⁺, CD4⁺CD25^high, CD4⁺CD25^high Foxp3⁺ T cells in the venous blood were detected in the two groups at 7 days after cell transplantation. Additionally, rat myocardial tissues were taken and observed pathologically. RESULTS AND CONCLUSION:The survival time of the cell transplantation group was significantly longer than that of the control group (P < 0.05). The expression level of interleukin-2 showed no significant difference between the two groups (P > 0.05), but the level of interleukin-10 in the cell transplantation group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the percentage of CD4⁺/CD8⁺, CD4⁺CD25^high, CD4⁺CD25^high Foxp3⁺ and CD4⁺ T cells was significantly higher, and the percentage of CD8⁺ T cells was significantly lower in the cell transplantation group (P < 0.05). Histopathological findings showed that there were a small amount of infiltrated lymphocytes in the cell transplantation group with the presence of slight bleeding and edema, and these inflammatory reactions were milder than those in the control group. These findings indicate that bone marrow mesenchymal stem cell transplantation can effectively reduce the rejection in rats undergoing heart transplantation.     

骨髓间充质干细胞在微损伤环境中修复骨不连

背景：骨髓干细胞能够增殖再生,与传统的手术治疗方案结合能明显增强骨不连的治疗效果,具有重要的应用价值。 目的：探讨骨髓间充质干细胞在微损伤环境中对骨不连的治疗效果。 方法：选取清洁级纯种新西兰大白兔40只,采用随机数字表法分为实验组与对照组,每组20只。按照手术操作流程获取胫骨骨髓,分离培养骨髓间充质干细胞,待细胞增殖到第3代够107数量级时,进行超顺磁氧化铁纳米粒子培养标记。在兔前肢桡骨中段约15 mm处造成骨缺损,骨缺损6周发生骨不连。实验组大白兔将骨髓间充质干细胞与髂骨碎粒一起植入骨缺损处。对照组不进行干细胞移植,于骨缺损处植入髂骨碎粒。术后12周内,观察大白兔骨不连部位大体形态、X射线片、病理学染色结果。 结果与结论：实验组术后明显发现骨痂,骨缺损处逐渐修复,直至完全愈合;对照组大白兔骨不连部位没有骨痂,骨髓腔封闭,充填肉芽组织。实验组有增生活跃的软骨组织,碎粒融合,骨缺损处出现类骨质,成骨细胞进行性增加;对照组软骨增生差,有大量死骨,骨粒未融合,没有成骨细胞。实验组桡骨缺损部位X射线显示云雾状影,骨髓腔恢复再通,骨骼塑形好;对照组骨碎粒吸收较少,骨髓腔部分闭塞,骨骼未连接,缺损处硬化。结果显示在局部微损伤环境中,骨髓间充质干细胞能够增殖分化为成骨细胞,修复骨缺损导致的骨不连具有显著效果。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

冠心病与非冠心病患者骨髓间充质干细胞基本生物学特性比较

BACKGROUND:It is unclear whether the body differences in patients with different types of heart diseases affect the characteristics and performance of stem cells. OBJECTIVE:To explore the biological characteristics of bone marrow mesenchymal stem cells from patients with different types of heart diseases. METHODS:Bone marrow mesenchymal stem cells were extracted using density gradient centrifugation from the bone marrow of 27 patients with coronary heart diseases and 20 patients with other heart diseases. Cell morphology was observed in the two groups. CD13, CD34, CD45, CD54, CD106 and CD44 positive expression was detected by flow cytometry. Cell proliferation was detected by MTT method, and the in vitro cell growth curves of the two groups were described. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cells of the two groups showed a long spindle shape, and there was no significant difference in the cell morphology between the two groups. In all the patients, the positive rates of CD34 and CD455 were less than 3.0%, while the positive rates of CD13 and CD44 were higher than 95.0%. However, the positive rates of CD54 and CD106 were higher in patients with coronary heart disease as compared with those with other heart diseases (P < 0.05). The in vitro growth curves of cells in the two groups were basically consistent, and the cell proliferation was only a little higher in the patients with other heart diseases compared with those with coronary heart disease. Experimental results show that different types of heart diseases in patients have no influence on morphology and proliferation of bone marrow mesenchymal stem cells, but some function-related proteins may exhibit certain difference in their expressions.     

高质量浓度肿瘤坏死因子α对骨髓间充质干细胞的损伤作用

BACKGROUND:Stem cell transplantation has achieved good results in the treatment of cerebral ischemia, and how to reduce apoptosis of transplanted cells has become the focus of the therapy. OBJECTIVE:To investigate the injured effect of tumor necrosis factor alpha (TNF-α) on bone marrow mesenchymal stem cells and its mechanism. METHODS:Primary cultured bone marrow mesenchymal stem cells from Sprague-Dawley rats were treated with 200 μg/L TNF-α for 6 hours. Cell vitality was assayed by MTT, and cell apoptosis was observed by Hoechst33342 staining. Apoptotic rate was detected by Annexin-V/PI double staining. Level of oxidative stress was evaluated by determination of malondialdehyde and superoxide dismutase levels. The protein expressions of phosphorylated-Akt, Akt, phosphorylated-FoxO1, FoxO1 were detected by western blot analysis. RESULTS AND CONCLUSION:After treatment with TNF-α, the cell vitality of bone marrow mesenchymal stem cells decreased, the apoptotic rate increased, and the cells were arrested in the S phase. Moreover, the oxidative stress level was elevated, and the protein expression of phosphorylated-Akt and phosphorylated-FoxO1 was significantly reduced compared with the control group (P < 0.05). These results suggest that TNF-α at high level contributes to the S-stage arrest, responsible for the apoptosis processes of bone marrow mesenchymal stem cells via the Akt-FoxO1 pathway.     

移植骨髓间充质干细胞肝癌大鼠细胞凋亡及炎症因子的动态变化

BACKGROUND:Bone marrow mesenchymal stem cell transplantation has not been thoroughly reported on its effects on apoptosis in hepatoma carcinoma cells and inflammatory factor level. OBJECTIVE:To investigate the effect of rat bone marrow mesenchymal stem cells on dynamic change of inflammatory factors and cell apoptosis during hepatocarcinogenesis. METHODS:Sixty healthy Sprague-Dawley rats were divided randomly into healthy group (n=30), control group (n=30) and transplantation group (n=30). Healthy group was given ordinary feed and normal water, while other groups were given diethylnitrosamine solution in drinking water to induce liver cancer models. Then, rats in the transplantation group were subjected to bone marrow mesenchymal stem cell transplantation via the tail vein. Two weeks after cell transplantation, CXCL5, interleukin-8 and interleukin-6 levels were tested by ELISA, mRNA level of hepatocyte nuclear factor 1α detected by RT-PCR, expression of Bcl-2 and Bax in liver tissue measured by immunohistochemical method, and liver cancer cell apoptosis index detected by TUNEL technique. RESULTS AND CONCLUSION:After modeling, the expressions of CXCL5, interleukin-8 and interleukin-6 in the control group were significantly higher than those in the healthy group (P < 0.05), while these indexes were reduced significantly after bone marrow mesenchymal stem cell transplantation (P < 0.05) and close to the normal levels (P > 0.05). Bone marrow mesenchymal stem cell transplantation significantly up-regulated the mRNA level of hepatocyte nuclear factor 1α in the liver tissue that was decreased obviously after modeling (P < 0.05). In addition, the expression of Bcl-2 was reduced, while the expression of Bax and the apoptosis index increased significantly in the transplantation group compared with the control group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cell transplantation contributes to hepatocyte differentiation and regeneration in liver cancer rats by reducing serum inflammatory factor levels and promoting apoptosis in hepatoma carcinoma cells.     

非接触共培养条件下骨髓间充质干细胞向类髓核细胞的诱导分化

BACKGROUND:Mesenchymal stem cells (MSCs) for the treatment of degenerative disc diseases present a new therapeutic strategy for the structural and functional recovery of the degenerative intervertebral disc. OBJECTIVE:To study the differentiation potential of rabbit bone marrow MSCs (BMSCs) into nucleus pulposus-like cells. METHODS:Passage 3 BMSCs and nucleus pulosus cells (NPCs) extracted from rabbits were assigned into simple BMSCs culture, simple NPCs culture or co-culture group. In the former two groups, BMSCs or NPCs were cultured in 6-well culture plates. In the co-culture group, passage 3 BMSCs and NPCs were cultured on the upper or lower layer of the 6-well Transwell chamber, respectively. Cell morphology was observed by inverted contrast phase microscope. At 7 days of culture, immunocytochemistry, RT-PCR and western blot were used to examine the expression levels of type II collagen and aggrecan. RESULTS AND CONCLUSION:At 7 days of co-culture, BMSCs were polygonal or short spindle-shaped, with no fiber-like changes, and cell morphology was close to that of NPCs. Additionally, the expression levels of type II collagen and aggrecan in the BMSCs co-cultured with NPCs were similar to those in the NPCs cultured alone, but significantly higher than those in the BMSCs culture only. Overall, these results show that coculture of BMSCs with NPCs can induce BMSCs differentiating into an NPCs phenotype, indicating that BMSCs may provide sufficient sources of cells for the biological treatment of degenerative disc diseases.