细胞移植治疗椎间盘退行性变的研究概况

目的 综述细胞移植治疗椎间盘退行性变疾病的研究现状. 方法 查阅近年来有关细胞移植治疗椎间盘退行性变疾病的国内外相关文献.进行回顾及综合分析. 结果 将椎间盘源性细胞或BMSCs通过单纯细胞移植、复合胶原蛋白支架移植等方法移植入退变椎间盘内,均能够表达类髓核细胞表型,增加ECM合成,缓解甚至抑制椎间盘进一步退变. 结论 细胞移植是目前治疗椎间盘退行性变疾病较为理想的方法.     

干细胞治疗椎间盘退变性疾病的研究进展

椎间盘退变性疾病(disc degeneration disease,DDD)是引起腰腿疼痛的主要病因之一.目前其治疗主要分非手术治疗和手术治疗两大类,这些治疗虽取得了一定疗效,但并不能从根本上延缓甚至逆转椎间盘的退变.随着对椎间盘退变研究的不断深入,发现椎间盘退变与髓核内代谢失衡、髓核细胞凋亡增加、细胞外基质(如蛋白多糖和Ⅱ型胶原)分泌减少有关.因此,如何增加椎间盘内的髓核细胞,促进细胞外基质的分泌,增加液体含量,改善髓核内微环境是治疗DDD的关键所在.近年来,研究人员利用干细胞治疗DDD发现干细胞有望从根本上延缓甚至逆转椎间盘的退变,目前已成为治疗DDD的研究热点之一.现就干细胞治疗DDD的相关研究进展综述如下.     

髓核细胞鉴定方法的相关研究进展

腰椎间盘突出症是引起腰痛的主要原因,目前治疗方法包括药物、手术等,这些方法不能阻止或延缓腰椎间盘突出症的病理进程,如腰椎融合术反而可能促进相邻椎间盘的退变病理进程.干细胞移植可以分化为髓核样细胞或/和可以促进退变椎间盘内髓核细胞的增殖,是目前研究的热点,但是,髓核细胞和关节软骨细胞表型非常相似,这些细胞均表达Ⅱ型胶原、SOX-9和聚集糖胺聚糖(目前最常用这三个指标来鉴定这些细胞),如何准确地区分干细胞是向髓核细胞方向分化还是向关节软骨细胞方向分化,这是研究椎间盘及关节疾病的基础,特别是我们如何准确高效地鉴定出髓核细胞是研究椎间盘退变的关键.现就如何准确高效地鉴定出髓核细胞综述如下.     

细胞移植治疗椎间盘退行性变研究进展

目前认为椎间盘退行性变是引起腰背痛的主要原因,也是骨科治疗的难点。临床上常采用椎管减压、腰椎融合或单纯髓核摘除手术治疗,可纠正脊柱不稳,改善临床症状,但无法使退变的椎间盘恢复正常的生理功能。应用细胞疗法治疗椎间盘退行性变,近年已成为生物学治疗领域的热点。多项研究显示,将自体椎间盘细胞或间充质干细胞作为种子细胞植入退变的椎间盘中,可有效地补充细胞数量,增加细胞外基质含量,从而延缓椎间盘退变进程,甚至修复受损的椎间盘。     

髓核细胞鉴定的研究进展

目前髓核细胞尚无特异性细胞标记,通常认为来源于髓核并表达Ⅱ型胶原、SOX-9和聚集糖胺聚糖的细胞即髓核细胞。仅以Ⅱ型胶原、SOX-9和聚集糖胺聚糖来在定义髓核细胞显然是不足的,因为上述细胞标记无法鉴别髓核细胞与软骨细胞。干细胞治疗椎间盘退行性疾病是目前的研究热点之一,精确鉴定髓核细胞在干细胞治疗椎间盘退变的背景下具有重要意义。     

骨髓间充质干细胞向髓核细胞的转化

骨髓间充质干细胞具有多向分化潜能,在不同环境下可诱导分化为多胚层来源的细胞,如:骨细胞、软骨细胞等.椎间盘退变主要源于髓核细胞及细胞外基质的变化,髓核细胞为类软骨细胞,随着细胞生物工程技术和分子生物学的进展,利用骨髓间充质干细胞结合细胞载体可再生出组织工程化的髓核细胞,将其植入椎间盘,从而阻止和逆转椎间盘退变,为治疗椎间盘退变提供一条新途径.     

渗透压及弹性增强结合蛋白在椎间盘退变中作用的研究进展

<正>随着中国人口的老龄化,颈椎病、腰椎椎间盘突出症等脊柱退行性相关疾病的发生率日趋升高,给人民群众造成了巨大的健康威胁和经济负担[1]。椎间盘退变(intervertebral disc degeneration,IDD)是导致脊柱退行性相关疾病的主要病理基础,其发生机制较为复杂,至今尚无明确定论。IDD病理特点主要包括:髓核细胞数量减少和功能下降,细胞外基质成分改变,如蛋白聚糖、Ⅱ型胶原等大分子的合成     

白细胞介素与椎间盘退变的研究进展

<正>在美国,腰背痛会影响到85%的人群,每年的医疗保健及相关费用大约为100亿美元,而椎间盘的退变是引起腰背痛的主要原因。椎间盘的退行性病变(intervertebral disc degeneration,IVDD)是指随着年龄的增长,髓核、纤维环及软骨终板等椎间盘各个组织的老化退变,是一系列脊柱退行性疾病的前提和病理基础。多数学者认为椎间盘的退变是多因素协同作用     

椎间盘源性干细胞的基础研究进展

正椎间盘退变(intervertebral disc degeneration,IVDD)是脊柱退行性疾病的重要病理基础之一。目前治疗椎间盘退变性疾病的方法主要集中于缓解症状,而非解决根本问题或恢复椎间盘本身的结构和功能[1]。为了修复重建椎间盘组织,研究者研发了许多方法并取得了一定成果,例如干细胞治疗[2]和基因治疗[3]等。其中,干细胞生物治疗一直是再生医学领域的研究热点。     

椎间盘组织工程种子细胞研究进展

椎间盘退行性变临床上常见,目前尚无有效治疗方法。椎间盘组织工程学技术以种子细胞为核心、支架材料为载体,为椎间盘退变治疗提供了新思路,其中种子细胞是最基本、最关键的环节。目前椎间盘组织工程研究常用种子细胞包括椎间盘细胞和干细胞。该文就椎间盘细胞中髓核细胞、纤维环细胞、脊索细胞,干细胞中骨髓间充质干细胞及其他干细胞在椎间盘组织工程种子细胞研究中的进展作一简要综述。     

Nestin-lineage cells contribute to the microvasculature but not endocrine cells of the islet

To clarify the lineage relationship between cells that express the neural stem cell marker nestin and endocrine cells of the pancreas, we analyzed offspring of a cross between mice carrying a nestin promoter/enhancer-driven cre-recombinase (Nestin-cre) and C57BL/6J-Gtrosa26(tm1Sor) mice that carry a loxP-disrupted beta-galactosidase gene (Rosa26). In nestin-cre(+/tg);R26R(loxP/+) embryos, cre-recombinase was detected in association with nestin-positive cells in the pancreatic mesenchyme with some of the nestin-positive cells lining vascular channels. In postnatal mice, pancreatic beta-galactosidase expression was restricted to vascular endothelial cells of the islet and a subset of cells in the muscularis of arteries in a distribution identical to endogenous nestin expression. Ex vivo explants of mouse pancreatic ducts grew dense cultures that costained for nestin and beta-galactosidase, demonstrating recombination in vitro. The cultures could be differentiated into complex stereotypic structures that contain nestin- and insulin-expressing cells. Nestin-cre(+/tg);R26R(loxP/+)-derived duct cultures showed that insulin-positive cells were negative for beta-galactosidase. These results indicate that both in vivo and in vitro pancreatic endocrine cells arise independently of nestin-positive precursors. The apparent vascular nature of the nestin-positive cell population and the close association with endocrine cells suggest that nestin-positive cells play an important role in the growth and maintenance of the islet.     

胰岛素样生长因子1基因转染对大鼠肌腱细胞生长的促进作用

目的　探讨外源性胰岛素样生长因子 1(IGF 1)基因转染对离体培养的肌腱细胞分裂增殖的促进作用。方法　用逆转录多聚酶链反应 (RT PCR)和酶联免疫吸附试验 (ELISA )检测转染 48h后细胞IGF 1mRNA和活性蛋白的表达 ;3 H TdR掺入法检测转染后 2 4、48、72h肌腱细胞DNA的合成情况 ;并用激光共聚焦显微镜对转染 48h后细胞Ⅰ型胶原的免疫荧光染色行定量分析。结果　IGF 1mRNA和活性蛋白在转染细胞内正确表达 ;实验组每分钟闪烁值 (CPM )为2 3 5 5 .75± 5 41.98,而对照组CPM值为 114 9.0 0± 485 .3 0 ,两者差异有显著性 (P <0 .0 5 ) ;转染组Ⅰ型胶原荧光强度较对照组明显增强 (转染组 76.2 0± 3 2 .2 3 ,对照组 3 8.84± 11.10 ,P <0 .0 1)。结论　外源性IGF 1基因转染能够促进肌腱细胞的分裂增殖和Ⅰ型胶原的分泌。     

丙戊酸钠增强免疫细胞对人膀胱癌细胞的杀伤作用

目的:探讨丙戊酸钠(VPA)对膀胱癌细胞MICA表达的影响以及所产生的肿瘤免疫作用,以期为防治膀胱癌复发和浸润提供新的治疗方法。方法:采用不同浓度VPA处理人膀胱尿路上皮癌T24细胞后,用半定量RT-PCR和流式细胞术检测癌细胞中MICA mRNA和蛋白表达。用乳酸脱氢酶法检测外周血单个核细胞(PBMCs)对经VPA处理的T24细胞的杀伤作用。结果:VPA从mRNA和蛋白水平诱导T24细胞表达MICA,并增强T24细胞对PBMCs细胞毒作用的敏感性。结论:VPA能增强膀胱癌对免疫细胞杀伤作用的敏感性,可作为防治膀胱癌的辅助药物。     

Dendritic cells exposed to nacystelyn are refractory to maturation and promote the emergence of alloreactive regulatory t cells

BACKGROUND: Drugs blocking dendritic cell (DC) maturation might be useful in transplantation by inhibiting the induction of primary alloimmune responses and promoting the emergence of regulatory T lymphocytes (Treg). We investigated the effects of Nacystelyn (NAL), an N-acetyl-L-cysteine derivative, on human DCs, paying attention to the T-cell responses elicited by NAL-treated DCs in vitro. METHODS: Lipopolysaccharide (LPS) was used to induce the maturation of DCs naturally present in blood or generated from human monocytes cultured in interleukin-4 and granulocyte-macrophage colony-stimulating activity. We first analyzed the consequences of NAL on cytokine production and expression of major histocompatibility complex class II and costimulatory molecules. Monocyte-derived DCs were then used as stimulators in mixed leukocyte cultures with naive CD4 T cells. Cytokine levels were measured in culture supernatants; the phenotype of T cells and their capacity to inhibit the proliferation of third-party T-cell responders was determined at the end of the culture. RESULTS: NAL proved to be a potent inhibitor of DC maturation in whole blood experiments and on monocyte-derived DCs. Alloreactive T cells stimulated with DCs pretreated with LPS in the presence of NAL produced much less interferon-gamma but similar levels of interleukin-13 compared with DCs treated with LPS alone. Immature DCs induced Treg, which was not observed with mature DCs. DCs cultured with LPS in the presence of NAL were as efficient as immature DCs to generate alloreactive T cells with regulatory activity. CONCLUSIONS: NAL is a potent inhibitor of DC maturation, which might be useful to promote allograft acceptance by inducing the differentiation of allospecific Treg.     

干细胞向生殖细胞分化的研究进展

干细胞(SCs)具有在体外分化为生殖细胞的潜能,为研究生殖细胞(GCs)早期发育提供了良好的模型,并将为干细胞移植修复生殖功能提供细胞资源。本文综述了胚胎干细胞/诱导多能干细胞(ESCs/iPSCs)、新生儿附属物来源干细胞(NDSCs)以及成体干细胞(ASCs)向生殖细胞分化所取得的研究进展,同时总结了各类干细胞向生殖细胞分化时所遇到的障碍及所面临的挑战,为干细胞在生殖医学领域的应用提供理论依据。     

Prepare cells to repair the heart: mesenchymal stem cells for the treatment of heart failure

Heart failure is one of the most important cardiovascular diseases, with high mortality, and invasive treatment such as mechanical circulatory support and cardiac transplantation is sometimes required for severe heart failure. Therefore, the development of less invasive and more effective therapeutic strategies is desired. Cell therapy is attracting growing interest as a new approach for the treatment of heart failure. As a cell source, various kinds of stem/progenitor cells such as bone marrow cells, endothelial progenitor cells, mesenchymal stem cells (MSC) and cardiac stem cells have been investigated for their efficacy and safety. Especially, bone marrow-derived MSC possess multipotency and can be easily expanded in culture, and are thus an attractive therapeutic tool for heart failure. Recent studies have revealed the underlying mechanisms of MSC in cardiac repair: MSC not only differentiate into specific cell types such as cardiomyocytes and vascular endothelial cells, but also secrete a variety of paracrine angiogenic and cytoprotective factors. It has also been suggested that endogenous MSC as well as exogenously transplanted MSC migrate and participate in cardiac repair. Based on these findings, several clinical trials have just been started to evaluate the safety and efficacy of MSC for the treatment of heart failure.     

小鼠iPS细胞具备诱导性原始生殖细胞分化潜能的研究

目的:探讨小鼠诱导性多能干细胞IP14D-1是否具备诱导性原始生殖细胞(induced primordial germcells,iPGCs)分化潜能,以及特异基因表达变化及可能机制。方法:未分化IP14D-1培养扩增,分化形成诱导性拟胚体(induced embryoid bodies,iEBs)。RT-PCR和免疫荧光分别检测4、7、9 d的iEBs中Lin28、Blimp1、Stra8和Mvh的表达变化和蛋白定位情况。结果:未分化IP14D-1同小鼠胚胎干细胞(mouse embryonic stem cells,mESCs)相同,Lin28表达较弱,Blimp1表达相对较强,Mvh和Stra8也在这两种细胞及其相应iEBs和EBs中表达,但均无明显差别。IP14D-1分化形成的iEBs从4 d生长到7 d时,Lin28表达逐渐增强,到9 d时表现为下降,Blimp1表达则随着iEB生长时间延长而逐渐降低。结论:建立了IP14D-1和相应拟胚体(iEBs)的完善、稳定的培养及分化体系;未分化IP14D-1与mESCs在Lin28、Blimp1、Mvh和Stra8表达方面无明显差别;iEB和EBs的Mvh和Stra8表达也无明显差别。IP14D-1及iEBs具有iPGCs分化潜能,且可能是7 d的iEBs内iPGCs分化数量较多,之后进入iPGCs分化的Lin28低表达时期。     

Elastase activity enhances the adhesion of neutrophil and cancer cells to vascular endothelial cells

BACKGROUND: Elastase activity in cancer cells has been reported to promote their metastasis. Hence, we analyzed the influence of elastase activity of cancer cells on their responsive adhesion to vascular endothelial cells. MATERIALS AND METHODS: Human pancreatic (S2-007, S2-013, S2-020, S2-028) and colonic (COLO205) cancer cell lines were used. S2-007, S2-013, and S2-020 possess high elastase activity, whereas S2-028 and COLO205 have low elastase activity. Adhesive reactions of these cancer cells and neutrophils to TNFalpha-activated HUVEC were analyzed. Bound cells onto HUVEC were counted after incubation for 10 min. The effects of suppression of elastase activity by ZD8321, a potent elastase inhibitor, and supplementation of human neutrophil elastase (NE) on the adhesive reactions were also analyzed. In addition, E-selectin expression on HUVEC and concentrations of soluble E-selectin in the medium were measured. RESULTS: Adhesion of cells with high intracellular elastase activity to TNFalpha-activated HUVEC was suppressed by ZD8321. On the other hand, adhesion of cells with low elastase activity was enhanced by exogenous NE. Expression of E-selectin, a key molecule in leukocyte-endothelial cell interaction, on HUVEC was increased by NE. Soluble E-selectin concentration in the medium increased after the adhesive reaction between neutrophils and HUVEC. This increase was thought to be due to the shedding of cell surface E-selectin. Such responses were inhibited by ZD8321. CONCLUSION: Elastase activity has a biological function of stimulating both the E-selectin expression on HUVEC and the resultant adhesive reaction of cancer cells with them. Inhibition of elastase activity is a potent strategy for controlling cancer metastasis.     

The potential of bone marrow-derived cells to differentiate to glomerular mesangial cells.

Bone marrow stem cells (BMC) develop into hematopoietic and mesenchymal lineages but have not been known to differentiate into glomerular cells. To investigate whether such differentiation is possible, a search was made for donor glomerular cells in lethally irradiated C57BL/6j (B6) mice given transplants of BMC from syngeneic mice transgenic for green fluorescence protein (GFP) ([GFP-->B6] mice). After the recipients of donor BMC manifested GFP-positive cells in their glomeruli, the numbers of such cells increased markedly, in a time-dependent manner, from 2 wk to 24 wk after bone marrow transplantation. Immunohistochemical analyses revealed that most GFP-positive cells in the glomeruli were neither macrophages nor T cells. With the use of a laser-scanning confocal microscope, GFP-positive cells were observed within the mesangium of [GFP-->B6] mice. Furthermore, indirect immunofluorescence assays demonstrated that desmin-positive cells in the glomeruli of these chimeric mice were also positive for GFP. Among glomerular cells isolated from [GFP-->B6] mice 24 wk after bone marrow transplantation and then cultured, the majority of cells (approximately 84%) stained for desmin and approximately 60% of the desmin-positive cells expressed GFP. In addition, these GFP-positive cells in the cultures contracted in response to angiotensin II stimulation. These results suggest that bone marrow-derived cells may have the potential to differentiate into glomerular mesangial cells.