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猪胎儿神经干细胞的分离培养和分化 总被引:1,自引:0,他引:1
本研究旨在从猪胎儿脑组织中分离培养神经干细胞,观察神经干细胞生长特性和体外增殖、分化特点.利用神经干细胞培养体系,从胎龄30 d的猪胎儿脑组织中分离培养神经干细胞并诱导神经干细胞贴壁分化,采用RT-PCR技术检测干细胞和分化细胞表面标志或相关基因.结果成功分离培养出神经干细胞,神经干细胞具有分化潜能.神经干细胞中Nestin表达强阳性,β-actin、DCX、Hesl、Oct4、Desmin、CD-90、Nanog和Sox2表达阳性;体外诱导的神经干细胞可以分化为星形胶质细胞(表达GFAP)、少突胶质细胞(表达GalC)和神经元细胞(表达NF、NSE和MAP2).结果提示,从猪胎儿脑组织分离神经干细胞具有可行性和有效性,神经干细胞具有自我更新、增殖和分化潜能. 相似文献
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为了解BMPs在SVZa神经干细胞增殖和分化过程中的作用,在使用不同浓度BMP4诱导SVZa神经干细胞增殖分化的基础上,利用启动子活体荧光标记技术,动态显示BMP4的表达.结果表明,低浓度(1~5ng/mL)BMP4促进sVZa神经干细胞的增殖,而高浓度BMP4(10~100ng/mL)抑制其增殖;BMP4在分化的早期(10d)促进神经元的分化,而在4d以后抑制神经元的分化;加入BMP4的拮抗剂Noggin可以阻断其作用,在嗅球,BMP4的主要作用可能是促使神经元祖细胞退出细胞周期启动分化,在RMS可能为促进其增殖并维持神经元祖细胞状态,而在SVZa,BMP4则主要促进神经干细胞向星形胶质细胞分化。 相似文献
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本研究旨在获得妊娠中期猪羊水来源千细胞,并通过用EGFP对干细胞进行标记,为以EGFP作为示踪标记对干细胞进行体内移植研究奠定基础.利用羊水来源干细胞培养技术体系,从胎龄60 d猪胎儿羊水中分离获得干细胞,通过脂质体介导转染将EGFP基因导入干细胞,诱导转基因干细胞向肌细胞和神经细胞分化,观察其分化特点.采用RT-PCR技术检测干细胞和分化细胞表面标志或相关基因.结果成功分离培养出妊娠中期猪羊水来源干细胞,并获得转EGFP基因干细胞.干细胞在表达EGFP的同时仍具有分化潜能.干细胞中Oct4、CD-90和Sox2表达阳性;体外诱导的干细胞能分化为肌细胞(表达myf-6和myoD)、星形胶质细胞(表达GFAP)、少突胶质细胞(表达GalC)和神经元细胞(表达NF、NSE和MAP2).研究表明,从妊娠中期猪胎儿羊水中分离干细胞具有可行性和有效性,转EGFP基因干细胞具有自我更新、增殖和分化潜能,可以用EGFP对羊水来源干细胞进行标记、追踪,为EGFP作为示踪标记对干细胞用于体内移植研究奠定了基础. 相似文献
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雄性成年昆明小鼠分为亚硝酸盐染毒组和生理盐水对照组,通过灌胃进行亚硝酸盐染毒,用BrdU腹腔注射和免疫组织化学方法研究齿状回神经干细胞的增殖和新生细胞的存活状况,用多重免疫荧光标记研究神经干细胞的分化和迁移方向。结果显示,染毒组小鼠齿状回颗粒层中的BrdU阳性细胞数量、存活率均极显著低于对照组(P<0.01),表明亚硝酸盐中毒对小鼠新生神经干细胞的增殖和存活产生了影响。染毒组BrdU/NeuN双阳性细胞的数量极显著低于对照组(P<0.01)。与对照组相比,染毒组中神经干细胞的迁移方向无显著性差异(P>0.05)。结果表明,亚硝酸盐中毒能显著抑制小鼠神经干细胞增殖,降低新生细胞的存活率。 相似文献
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本研究旨在探究荷斯坦奶牛乳腺干细胞的多向分化潜能及催乳素对其增殖活性的影响。采用尼氏染色法对乳腺干细胞向神经细胞分化的情况进行鉴定,利用RT-PCR技术对诱导后的乳腺干细胞进行基因水平检测;MTT法检测催乳素对乳腺干细胞增殖活性的影响。结果表明:荷斯坦奶牛乳腺干细胞经成神经诱导后,细胞中有类似神经元的细胞出现,并且细胞周围出现许多微管样的结构,经尼氏染色呈阳性。诱导后神经细胞标记基因NSE和β-Tubulin Ⅲ呈阳性表达。催乳素刺激可促进乳腺干细胞的增殖,低浓度时乳腺干细胞的活力较好,且100ng·mL~(-1)为增殖最适浓度。 相似文献
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为探讨原代Wistar大鼠海马神经细胞的体外培养方法,本研究取新生24 h内的Wistar大鼠海马组织,无菌剪碎后采用胰酶消化法分离细胞,用含20%胎牛血清的DMEM/F12培养基培养,逐日在倒置相差显微镜下观察。结果发现,从海马组织中分离出的神经细胞具有增殖能力,细胞对数生长期为2~8 d,最长培养30 d;细胞经免疫荧光鉴定Nestin表达呈阳性;免疫组织化学结果显示,在传代培养细胞的胞体和突起均有NF阳性标记物,GFAP抗体和CD68抗体显色均为阴性。由此可见,分离培养的细胞是具有自我更新增殖和多分化潜能的神经元细胞。 相似文献
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昆虫的神经系统属于腹神经索型,控制着激素分泌、进食、运动以及支配内脏器官的活动,与脊椎动物的神经系统相比较,其结构简单易于实验操作。对昆虫神经系统的研究,可发现特异性靶标细胞,用于开发新型环保杀虫剂等。此外,昆虫神经干细胞的分裂、分化调控机制与脊椎动物甚至人类有很多的相似性,因而对昆虫神经干细胞的研究可为人类退化性神经疾病研究提供借鉴。本文着重阐述昆虫特别是果蝇的神经系统结构,神经细胞的类型,成神经细胞(neuroblasts,NBs)以及神经干细胞分裂、分化调控机制等方面的研究进展,期望能为开展家蚕神经干细胞的研究提供参考。 相似文献
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Reasons for performing study: Two studies report variability in proliferation and limited adipocyte differentiation of equine peripheral blood‐derived adult mesenchymal stem cells, thus casting doubt on their adipogenic potential. Peripheral blood can be a valuable source of adult mesenchymal stem cells if cell culture conditions permissive for their adherence, proliferation and differentiation are defined. Hyperbaric oxygen treatment has been reported to mobilise haematopoietic progenitor stem cells into the peripheral blood in humans and mice, but similar experiments have not been done in horses. Objectives: To optimise cell culture conditions for isolation, propagation and differentiation of adult stem cells from peripheral blood and to assess the effect of hyperbaric oxygen treatment on adult stem cell concentrations. Methods: Peripheral blood was collected from the jugular vein of 6 research mares, and mononuclear cells were isolated. They were subjected to cell culture conditions that promote the adherence and proliferation of adult stem cells. The cells were characterised by their adherence, expression of cellular antigen markers, and trans‐differentiation. Each horse was subjected to 3 hyperbaric oxygen treatments, and stem cells were compared before and after treatments. Stem cells derived from adipose tissue were used as controls. Results: One‐third of the horses yielded viable stem cells from peripheral blood, positive for CD51, CD90 and CD105, and demonstrated osteocyte, chondrocyte and adipocyte differentiation. Hyperbaric oxygen treatment resulted in a significant increase in CD90‐positive cells. Horses that did not yield any cells pretreatment did so only after 3 hyperbaric oxygen treatments. Conclusions and potential relevance: Peripheral blood can be a valuable source of adult stem cells, if one can identify reliable equine‐specific markers, provide methods to increase the number of circulating progenitor cells and optimise cell culture conditions for growth and viability. Our findings are important for further studies towards technological advances in basic and clinical equine regenerative medicine. 相似文献
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Role of gap junctional intercellular communication (GJIC) through p38 and ERK1/2 pathway in the differentiation of rat neuronal stem cells 总被引:4,自引:0,他引:4
Yang SR Cho SD Ahn NS Jung JW Park JS Jo EH Hwang JW Jung JY Kim TY Yoon BS Lee BH Kang KS Lee YS 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(3):291-294
Gap junctional intercellular communications (GJIC) contributes to neural function in development and differentiation of CNS. In this study, we have investigated the expression of GJIC during the differentiation of neuronal stem cells and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced neuronal stem cell-derived cells from rat brain. During neuronal stem cell differentiation, expressions of Cx43 and 32 were increased for the duration of 72 hr, however the effect were decreased on the 7d. In the neuronal stem cell-derived cells, pretreatments with p38 MAP kinase inhibitor, SB203580, and MEK inhibitor, PD98059, could protect GJIC against TPA-induced inhibition of GJIC. Our data suggest that GJIC plays an important role during neuronal stem cell differentiation, and ERK1/2 and p38 MAP kinase signaling pathway may be closely related functionally to regulate gap junction in rat neuronal stem cell-derived cells. 相似文献
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K S Kang J E Park D Y Ryu Y S Lee 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(11):1183-1190
Endocrine disrupters are exogenous compounds thought to mimic the action of estrogen or other hormones and influence endocrine activity in the body (Juberg, 2000). These chemicals have adverse effects not only in the reproductive system but also in the central nervous system during development and throughout life. Polychlorinated biphenyls (PCBs) are a class of environmentally persistent and widespread halogenated hydrocarbons. It has been reported that PCBs are potential neurotoxicants. Endosulfan is an organochlorine insecticide that is extensively used to control pests in vegetables, cotton, and fruits. To determine the effect of 2, 2', 4, 4', 5, 5',-hexachlorobiphenyl(2, 4, 5-HCB) and endosulfan on embryo nervous system, we isolated neural stem cells from rat brain at embryonic day 17. Isolated neural stem cells showed pluripotenty. Stem cells could differentiate into neurons and glia. Neurite formation in endosulfan and 2, 4, 5-HCB treated cells. And it appeared to be decreased as compared with that in untreated cells. In order to know the neuro-toxic mechanisms of 2, 4, 5-HCB and endosulfan in neuronal stem cells, we investigated mitogen-activated protein kinase activity (MAPK) and gap junctional intercellular communication (GJIC). Endosulfan decreased the MAPK activity in dose dependent manner. Endosulfan and 2, 4, 5-HCB inhibited GJIC compared to the untreated cell by scrape loading dye transfer (SL/DT). 2, 4, 5-HCB and endosulfan decreased the expression of connexin 43 in dose dependent manner. These results indicated that 2, 4, 5-HCB and endosulfan may inhibit differentiation and proliferation of neural stem cells and gap junctional intercellular communication which play a crucial role in the maintenance of cellular homeostasis. 相似文献
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Isolation and culture of rabbit primordial germ cells 总被引:2,自引:0,他引:2
Kakegawa R Teramura T Takehara T Anzai M Mitani T Matsumoto K Saeki K Sagawa N Fukuda K Hosoi Y 《The Journal of reproduction and development》2008,54(5):352-357
Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells. 相似文献
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鸡胚胎干细胞是一种多能性干细胞,从X期胚盘分离胚盘细胞或早期鸡胚的生殖嵴分离原始生殖细胞,经体外长期抑制分化培养可得到鸡胚胎干细胞。为维持细胞在培养过程中的未分化状态,需要采用饲养层细胞培养,同时设计合理的培养液配方并添加多种抑制分化或促进增殖的细胞因子。通过碱性磷酸酶活性检测、胚胎表面特异性抗原检测、分化试验及嵌合体试验等方法,可对鸡胚胎干细胞进行准确鉴定。文章主要就鸡胚胎干细胞的分离、培养与鉴定方法的研究进展及其应用前景进行简要综述,为进一步发展更高效的鸡胚胎干细胞培养体系并应用于生产实践提供一定的借鉴。 相似文献
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