共查询到20条相似文献，搜索用时 15 毫秒
1.
AIM: To investigate the effect of cellular repressor of E1A-stimulated genes (CREG) on the spontaneous apoptosis of murine embryonic stem cells(ESCs)-derived embryoid bodies (EBs).METHODS: The murine ESCs R1 were infected with pDS-shRNA-CREG and pDS-shRNA-GFP retrovirus, respectively. R1, R1-GFP and R1-shCREG were cultured on STO feeder cells in DMEM supplied with leukemia inhibitory factor(LIF). Alkaline phosphatase(AKP) staining and teratoma formation assay inoculated into mouse myocardium were used to detect stemness of transfected ESCs. R1/EB, R1-GFP/EB and R1-shCREG/EB were produced by liquid suspension method. The expression of CREG and cleaved caspase-3 were analyzed by Western blotting and quantitative RT-PCR on day 7. The apoptotic rates of the 3 kinds of EBs were analyzed by flow cytometry(FCM) analysis with Annexin V/PI dual staining. RESULTS: The stably-transfected ES cells (R1-shCREG and R1-GFP) were obtained by screening the G418-resistant clones. R1-GFP and R1-shCREG had the stem cell properties similar to those of R1 detected by AKP staining. However, it was found that R1-shCREG didn't show the same almighty differentiation function as of R1 and R1-GFP by tumor experiments in mouse myocardium that it couldn't form teratomas analogue and had the lower ability of cell differentiation. Observation under phase-contrast microscope showed that the cell differentiation was inhibited while the number of cell death was increased in R1-shCREG/EB group. Compared with R1/EB and R1-GFP/EB, Western blotting analysis demonstrated that the protein expression of CREG was decreased to (78.0±1.3)% and (84.0±2.4)% on day 7, respectively. The mRNA expression of CREG was also decreased, but the expression of cleaved caspase-3 at the mRNA and protein levels was increased obviously. Annexin V/PI FCM assay indicated that the apoptotic rate of R1-shCREG/EB was significantly higher those that in other 2 groups on day 7. CONCLUSION: The down-regulation of CREG inhibits ESCs differentiation and promotes cell apoptosis. 相似文献
2.
AIM: To investigate the potential of murine epidermal stem cell (ESC) differentiation after seeded in a biodegradable carrier and implanted subcutaneously into syngeneic recipient mice. METHODS: ES cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a polyglycolic acid (PGA) net containing collagen gel, functioning as a cell carrier, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. RESULTS: The ESCs kept alive in the implant when observed under a fluorescent microscopy 3 weeks or longer after implantation, and could differentiate into hair follicle-like structure, glandular structure, and gave rise to additional structures displaying features resembling native dermis. No apparent rejection or severe side effects were observed at least 10 weeks post-implantation. CONCLUSION: It is feasible to use these ESCs as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages. 相似文献
3.
AIM: To investigate the differentiation of murine embryonic stem cells (ESCs) into hematopoietic stem cells (HSCs) by the supportive effects of human aorta-gonad-mesonephros (AGM) region and fetal liver (FL) stromal cells.METHODS: E14 ESCs were induced into embryoid body (EB) first. Then the cells from EB were further co-cultured with human AGM region and FL stromal cells in non-contact system. On day 6, the cells derived from EB were collected for Sca-1+c-Kit+ cell analysis by flow cytometry, colony forming unit (CFU) assay and teratoma formation checking. BALB/c female mice conditioned with lethal dose of γ-ray irradiation were transplanted with EB cells from different culture systems. The survival rates, engraftment of donor cells, reconstitution of hematopoietic were monitored.RESULTS: Sca-1+c-Kit+ cells in EB cells co-cultured with human AGM region and FL stromal cells had the value of (21.96±2.54)%, and the total CFU was as (520±52)/105 cells, which were statistically greater than those in EB cells only cultured with human AGM region stromal cells (P＜0.05). No teratoma was found in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region and FL stromal cells. In BALB/c female mice transplanted of EB cells co-cultured with human AGM region and FL stromal cells, the survival rate was 77.8%, and the peripheral blood cell count was obviously improved on day 14. PCR results showed the recipients all had sry gene copies from donor in bone marrow. The recipient mice transplanted with EB cells only cultured with human AGM region stromal cells all died within 15 days.CONCLUSION: Stromal cells from human AGM region and FL enhance the directed differentiation of ESCs into HSCs which can reconstruct hematopoiesis in vivo. 相似文献
4.
ZHANG Xu-chao CHEN Hui-qin HUANG Shao-liang WU Bei-yan HUANG Yong-lan CAI Yun 《园艺学报》2007,23(9):1747-1751
AIM: To direct embryonic stem cells (ESCs) into hematopoietic stem cells (HSCs) in vitro by simulating the hematogenic microenvironment in human early embryonic aorta-gonad-mesonephero (AGM) region.METHODS: Murine E14 embronic stem cell line was used for two-step differentiation.In the first step of primary differentiation,E14 ESCs were seeded into semisolid methylcellulose-based medium containing bone morphogenesis protein 4 (BMP4) and vascular endothelial growth factor (VEGF) for embryoid body (EB) formation.On days 3,6,9,12 and 15,single EB cells were analyzed for Flk-1+ cells amount through flow cytometry.In the second step,single cell from EB containing most Flk-1+ cells was further co-cultured with human AGM stromal cells in non-contact system.On co-culture days of 3,6,9 and 12 days,cells were collected for cell count,flow cytometry for Sca-1+c-kit+ cells analysis,and colony forming cell assay.RESULTS: During the EB formation,BMP4+VEGF promoted Flk-1+ cell genesis on day 9 at peak pencentage value of 27.53%±2.84％,which was statistically higher than that in control group as 8.77±1.10 (P<0.05).Collagenase-disassociated single cell from day 9 EB was co-cultured with human AGM stromal cells of hAGMS3 or hAGMS4 for further hematopoietic differentiation.On day 6 Sca-1+c-kit+ cells got to peak value as 7.31%±1.21％ ［(2.57±0.48) folds］ and 7.62%±1.52％ ［(2.35±0.36) folds］ in hAGMS3 and hAGMS4 feeder systems,respectively,both of which were greater than those values of no-stroma groups at the same culture duration (P<0.05).Colonogenic cell assay showed that these Sca-1+c-kit+ cells had ability of forming multiple lineage hematopoietic colonies.CONCLUSION: BMP4 in combination with VEGF promotes Flk-1+ cell genesis during EB formation in vitro.Stromal cells from early human embryonic AGM region further enhance the directed differentiation of these primitive cells into HSCs.This two-step induction differentiation model can be used for molecular mechanism study of ESCs hematopoietic differentiation. 相似文献
5.
AIM：To compare the effects of three different cell culture protocols：embryonic body (EB) formation，EB formation-monolayer and monolayer on differentiation of mouse embryonic stem (ES) cells into insulin-secreting cells.METHODS：E14.1 mouse ES cells were treated with GLP-1,betacellulin，activin A,bFGF and nicotinamide by using EB formation,EB formation-monolayer and monolayer culture protocol respectively for 30 days,then insulin expression was examined by RT-PCR,DTZ-staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flow cytometry.RESULTS：DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed in the differentiated cells for all the three groups.mRNAs of insulin and some other islet-related genes were detected,insulin expression was the strongest in EB formation-monolayer,and the weakest was in monolayer.The percentage of insulin-positive cells of the differentiated cells in the EB formation-monolayer group was higher than that in the EB formation group (P＜0.01),the latter was higher than that in the monolayer group (P＜0.01).CONCLUSION：Among the three cell culture protocols,EB formation-monolayer is the most effective approach in the induction of mouse ES cells to differentiate into insulin-secreting cells. 相似文献
6.
AIM:To investigate whether efficient production of neuron-like cells from embryonic stem cells (ESCs) can be indued by astrocyte-conditioned medium (ACM) in vitro. METHODS:Based on the 4-/4+ protocol established by Bain, two groups were studied:ATRA group, ATRA with ACM group. ESCs were induced into neuron-like cells by means of three-step differentiation in vitro. The totipotency of ESCs was identified by the observation of cells’ morphology and the formations of teratoma in immunocomprised mice. The cell differentiation was evaluated continuously by detection of the cellular specific markers of neural stem cell, neurons and astrocytes such as nestin, NF-200, NSE and GFAP using immuno-histochemistry assay. RESULTS:(1) The ESC-D3 cells kept the ability of differentiation into cellular derivations of all three primary germ layers after continuous passage culture. (2) The ratio of NF-200 and NSE positive cells in the cells induced by ATRA with ACM was higher than that in the cells induced by ATRA only. (3) Finally, the positive rate of the neuron-like cells was up to 73.5% in the group induced by ATRA with ACM. CONCLUSION:The ESCs are induced into neuron-like cells with high purity and efficiency by ATRA with ACM. 相似文献
7.
甲基胺草磷诱导大蒜多倍体的研究 总被引：2，自引：1，他引：1
以通过生理休眠的紫皮大蒜Z-006-10鳞茎茎尖为试材,利用固体培养基中添加甲基胺草磷的离体诱导方法,研究甲基胺草磷不同处理浓度和时间对紫皮大蒜体细胞染色体的诱导效果,并利用形态观察、根尖细胞染色体计数及流式细胞仪分析等方法对得到的再生植株进行倍性鉴定。试验结果表明：10 μmol?L-1甲基胺草磷处理6 d,四倍体诱导率最高,达32.87%。形态观察结果表明,得到的四倍体植株多倍体形态特征明显,与二倍体植株差异明显。根尖染色体压片观察表明,四倍体染色体数为2n=4x=32,对照为2n=2x=16。流式细胞仪倍性分析表明,四倍体体细胞DNA的相对含量是二倍体的2倍,其结果与染色体计数法鉴定结果一致。 相似文献
8.
LUO Wei LAI Ke-fang CHEN Ru-chong LIU Chun-li ZENG Yun-xiang HE Xin-ming ZHONG Shu-qing HE Meng-zhang LI De-rong ZHONG Nan-shan 《园艺学报》2006,22(5):943-947
AIM: To explore the pathological features of airway inflammation in patients with eosinophilic bronchitis (EB) and compared to those with cough variant asthma (CVA). METHODS: Flexible fibre optic bronchoscopy was performed in 11 patients with EB, 10 with CVA, 14 with bronchial asthma and 10 normal controls. The mean thickness of the basement membrane was measured by light microscopy. Using immunohistochemical and special staining, the localization and density of inflammatory cells (eosinophils, mast cells, T lymphocytes) were detected in bronchial submucosa in EB and CVA patients. RESULTS: The mean thickness of the basement membrane was significantly increased in the subjects with EB ［2.92 μm (2.10-6.50 μm)］, CVA ［5.64 μm (3.23-8.48 μm)］ and bronchial asthma ［9.08 μm (6.61-11.99 μm)］ rather than that in the normal controls ［2.08 μm (1.62-3.40 μm)］. There were also significant differences among the three groups. The number of mast cells and eosinophils in the bronchial submucosal from subjects with EB ［75 cells/mm2 (35-112 cells/mm2), 7 cells/mm2 (0-31 cells/mm2)］ was substantially decreased than those in subjects with CVA ［148 cells/mm2 (34-200 cells/mm2), 114 cells/mm2 (1-768 cells/mm2); P<0.05］. There was no significantly difference in T lymphocyte counts between the EB and CVA. CONCLUSIONS: EB is an inflammatory disorder of the airways with the characteristics of various inflammatory cell (eosinophils, mast cells and T lymphocytes) infiltration. The mean thickness of the basement membrane is less severe than that in CVA and bronchial asthma and the level of infiltration of inflammatory cells is less than that in CVA, which may be one of the reasons that airway hyperresponsiveness is rarely seen in EB. 相似文献
9.
多粘类芽孢杆菌CP7对荔枝霜疫霉菌的抗菌活性及其作用机制 总被引：4，自引：0，他引：4
多粘类芽孢杆菌Paenibacillus polymyxa CP7菌株分泌的抗菌物质对荔枝霜疫霉菌等真菌具有很强的抑杀作用。用光学显微镜、扫描电镜和透射电镜观察发现,荔枝霜疫霉菌菌丝体经CP7菌株的抗菌物质处理后, 菌丝发生不规则扭曲、萎蔫、干瘪,菌丝端部膨大等形态变化;经处理的菌体细胞组织结构受损,细胞膜结构模糊不清,看不到完整的外膜结构,细胞器消溶。处理后的荔枝霜疫霉菌孢子囊出现凹陷、干瘪、畸形;孢子囊外壁严重受损,原来厚实的外壁只剩一薄层,孢子囊内原有的微细结构消失,结构模糊不清,细胞器溶解,孢囊内物质大量外泄。抗菌物质还引起菌体细胞内磷的外泄,并对细胞呼吸速率和NADH氧化酶活力有抑制作用。 相似文献
10.
AIM: To establish hybrid mouse embryonic stem (ES) cell line from blastocysts of the (C57BL/6J×129/J) F1 mouse. METHODS: 3.5 days post-coitus (d.p.c.) blastocysts were cultured on mouse embryonic fibroblasts (MEFs) in the medium, after 3-4 days, Inner cell mass were picked up and disaggregated, then reseeded. After the ES-like colonies appeared, passaged them to give permanent ES cell lines. The pluripotent properties of ES cells obtained were analyzed by alkaline phosphatase (AKP) activity, expression of SSEA-1 and Oct-4, and their capacity to form teratoma. RESULTS: Two hybrid ES cell lines, SC1001, SC1002 were obtained from blastocysts of the (C57BL/6J×129/J) F1 genotype. Most of these ES cells had a normal karyotype and an XY sex chromosome composition. The pluripotent properties of the cell lines were analyzed on the basis of their alkaline phosphatase activity, expression of SSEA-1 and Oct-4, and their capacity to form teratoma in severe combined immunodeficiency (SCID) mice. CONCLUSION: Two hybrid mouse ES cell lines having pluripotent properties and capacity for long-term self renewal were generated from blastocysts of the (C57BL/6J×129/J) F1 genotype. 相似文献
11.
12.
以茶子属（Ｒｉｂｅｓ）的杂种植株为试材，对其核型和显带技术作了详细的研究。结果表明，染色体的风油精制备法比去壁低渗法和常规压片法能得分散良好、结构分明的核型图，并且未经Ｇ带处理也能够显示出Ｇ带，说明该方法可成为一种新的预处理药物应用于植物，特别像果树这类染色体数目多、体积小的染色体标本制备或Ｇ带的诱导。关键词 相似文献
13.
HE Yue GAO Yun-fang ZHANG Shao-ying ZHANG Yang-mei SUN Xiao-yong WANG Hui-ping 《园艺学报》2012,28(8):1500-1503
AIM:To investigate the effects of disuse and reloading on the calpain activity and cell membrane permeability of soleus muscle fibers in rats. METHODS:Tail-suspension(TS) rats were used as a muscular atrophy model. The female rats were randomly divided into control group(CON), control group with training(CON+T), 2-week TS group(TS) and 2-week TS with training group(TS+T). The changes of cell membrane permeability and calpain activity were detected with Evans blue(EB) as a tracer and casein zymography, respectively. RESULTS:The results showed that the EB was full of intercellular space in soleus muscle fibers in CON group, a few in TS group and much more in TS+T group, respectively. In TS group and TS+T group, some self-excitation bands appeared and absorbance of calpain hydrolysis bands was significantly increased compared with those in the controls. CONCLUSION:The results suggest that the cell membrane permeability in TS+T group raises and the membrane barrier function is damaged. This may attribute to the hydrolysis of membrane skeleton proteins by calpain. Calpain-mediated accelerated degradation of cytoskeleton proteins may be an important mechanism of skeletal muscle injury induced by reloading. 相似文献
14.
WANG Ji-liang LIN Jun-wen SHI Ze-jin TAI Ying-jie REN Jie HE Yi-gang HUANG Hua-yuan HE Shi-yong 《园艺学报》2007,23(3):509-513
AIM:To investigate the effect of silymarin on homocysteine-induced cell viability and apoptosis in human umbilical vein endothelial cells (HUVECs). METHODS:Cell viability was analyzed by using MTT and LDH assay. Apoptotic cells were detected by using DNA fragmentation and flow cytometric analysis. The level of intracellular reactive oxygen species (ROS) and the potential of mitochondrial membrane were determined by flow cytometric assay. The activity of caspase-3, -6 and -9 were measured with microplate spectrofluorometer. Protein levels were examined by Western blotting. RESULTS:Treatment of cultured HUVECs with HCY for 48 h induced a significant decrease in cell viability, and the percentage of apoptosis increased to 76.8%. The level of intracellular ROS and activity of caspase-3, -6 and -9 enhanced, and the red/green ratios of mitochondrial membrane decreased. However, simultaneous treatment with silymarin exhibited cytoprotective effects, reduced formation of the DNA ladder, prevented the levels of Bax and Bcl-2 proteins and the accumulation of ROS as well as caspase-3, -6 and -9 activation, reconverted the potential of mitochondrial membrane, and the percentage of apoptosis/necrosis was significantly decreased to 12.7% in a dose-dependent manner. CONCLUSION:These results demonstrate that silymarin has the protective capacity to antagonize HCY-induced apoptosis in HUVECs. The antiapoptotic action of silymarin may be partially dependent on an anti-oxidative stress effects, inhibition of caspases activity, and maintenance of mitochondria function. 相似文献
15.
DU Hui-bo SONG Wen ZHANG Li-min XING Li-qiang ZHANG Hui ZHAO Zi-gang NIU Chun-yu 《园艺学报》2014,30(4):686-692
AIM: To observe the effects of normal mesenteric lymph (NML) on the lung, heart and liver injuries and the phosphorylation levels of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK) in the mice with endotoxic shock (ES). METHODS: The NML was drained form health male BALB/c mice for the intervention of ES after the removal of cellular constituent. Lipopolysaccharide (LPS, 35 mg/kg) was intraperitoneally injected into the mice for the establishment of ES model. After 60 min of LPS injection, the administration of NML (1/15 of whole blood volume) was performed through the femoral artery in NML+ES group. Meanwhile, the mean arterial pressure (MAP) was monitored during the experiment. At 6 h after intraperitoneal injection of LPS or the corresponding time point, blood samples were harvested from the heart through apical centesis for determination of the biochemical indexes to reflect myocardial and hepatocyte injuries. Simultaneously, the lung, heart and liver tissue specimens from a fixed location were harvested for the observation of histomorphology and the measurement of phosphorylation levels of p38 MAPK, ERK1/2 and JNK. RESULTS: Compared with sham shock (SS) group, MAP in ES group and NML+ES group remarkably decreased at multiple time points after intraperitoneal injection of LPS. However, MAP in NML+ES group at 80 min, 90 min, 190 min, 210 min, 240 min, 250 min, 340 min, 350 min, and 360 min were significantly increased compared with ES group. There were normal structures in the lung, liver and myocardium of the mice in SS group, while the morphological damages of these tissues appeared in ES group. Meanwhile, the damages were attenuated in the mice of NML+ES group. The activities of AST, ALT and CK-MB in the plasma in ES group were remarkably higher than those in SS group. The CK-MB activity in NML+ES group was also increased compared with SS group, and the activities of AST and LDH-1 were lower than those in ES group. At 6 h after LPS injection, the phosphorylation levels of p38 MAPK, ERK1/2 and JNK in the lung tissues were remarkably increased. Meanwhile, no statistical difference of these indexes between the myocardial and hepatic tissues was observed. NML intervention decreased the phosphorylation levels of p38 MAPK in the lung tissues, and p38 MAPK, ERK1/2 and JNK in the myocardial tissues. CONCLUSION: The NML administration alleviates multi-organ injuries and reduces the phosphorylation level of p38 MAPK in the lung tissues in the mice subjected to ES. 相似文献
16.
AIM: To observe whether arachidonic acid (AA) could induce apoptosis in mouse fibroblast cell line L929 and the potential mechanism involved. METHODS: The viability and damaged degree of L929 was monitored by MTT and the release of lactate dehydrogenase (LDH). Lipid peroxidation in L929 was measured as malondialdehyde (MDA) content by colorimetric assay. Hoechst 33258 staining was used to observe AA-induced morphological changes. Agarose gel electrophoresis was used to detect DNA fragmentation. RESULTS: Treatment of L929 cell with AA for 24 h, in the range of 40-160 μmol/L, caused a great decrease in cell survival and increased MDA contents and the release of LDH simultaneously (P＜0.01). Cells treated by 160 μmol/L AA showed typical morphological changes of apoptosis. A "ladder" pattern representing fragmentation of DNA into oligonucleosome length fragments was observed 24 h after AA treatment. CONCLUSION: Higher concentration of arachidonic acid (80-160 μmol/L) induced apoptosis in mouse fibroblast cell line L929. The mechanism of its action might be related to lipid peroxidation. 相似文献
17.
AIM:To explore the mechanism and significance of the intestinal epithelial cellular membrane damage following burn serum. METHODS:The intestinal epithelial cell(IEC-6) were cultured. The changes of total membranous phospholipid contents fluidity of the IEC membrane were dynamically examined with fluorescence polarization technique and HPCE. RESULTS:In the early stage after stimulation by 20% burn serum, the membranous fluidity obviously decreased. The total phospholipid contents decreased, the content of PLA2 markedly increased. CONCLUSION:The serial changes in IEC after burned could result in the damages of IEC membrane structure, the integrity of cell membrane and function. 相似文献
18.
AIM: To study apoptotic injury induced by reactive oxygen species-hydrogen peroxide (H2O2) on cardiac myocytes.METHODS:Cultured rat neonatal cardiac myocytes were treated with H2O2 of various concentration to observe apoptotic injury of cardiomyocytes by agarose gel electrophoresis, Giemsa-stained smears of cell, and flow cytometry, meanwhile lactate dehydrogenas (LDH) and malondialdehyde(MDA) were determined to assess the effect of H2O2 on lipid peroxidation and permeability of the plasma membrane. RESULTS: 5 mmol/L H2O2 caused cultured cardiomyocytes apoptotic morphological characteristics, including nucleosomal DNA fragmentation in myocytes by agarose gel electrophoresis (DNA ladder), cell shrinkage, nuclear condensation, and chromatin margin by Giemsa-stained cell smears, and aneuploid peak(AP)-apoptotic bodies occurrence by flow cytometry.CONCLUSIONS: H2O2-induced apoptosis in myocytes was a time-and concentration-dependent process. Treatment with low concentration of H2O2(＜1 mmol/L) only caused cardiomyocyted early biochemical changes, such as increase of free radicals level and membrane permeability ,which were pro-apoptotic injurious features. High concentration of H2O2 (＞10 mmol/L) rapidly induced a necrotic form of death characterized by smeared patterns of DNA digestion on agarose gel electrophoresis and lethal membrane disruption (as measured by LDH release). Exposure of 5~10 mmol/L H2O2 induced cardiomyocytes apoptosis concurrently with biochemical changes of LDH and MDA increase in the medium. 相似文献
19.
AIM: To construct a knock-out vector for βmaj & βmin gene of β-globin gene and establish embryonic stem cell lines by gene knock-out in ES level for generating gene knock-out animal model. METHODS: Two homologous sequences 1.7 kb and 7.0 kb were inserted into basic plasmic vector pNTK, which contains positive and negative selection system of the neomycin resistance (Neo) and herpes simplex virus thymidine kinase (TK) resistant gene to construct a gene knock-out recombinant plasmic vector nominated β-pNTK, which was certified by polymerase chain reaction, enzyme digested and DNA sequencing. The β-pNTK that contains two homologous sequence was linearized with Sal I and introduced into the D3 line of ES cells by electroporation. Positive cloning of ES cells was selected with positive and negative selection system of G418 600 mg/L and 2.5 mol/L Gancyclovir for four weeks. ES cells clone were picked, explanded and certified by polymerase chain reaction, Southern blotting. Examination of the undifferentiated state and pluripotential characteristics of the cell lines were made with the alkaline phosphatase (AKP) staining and embryoid bodies formation test and teratomas formation in nude mice. RESULTS: A knock-out recombinant plasmic vector for βmaj & βmin gene of β-globin gene, β-pNTK, was constructed in C129 mouse, which contains 8.7 kb length inserted homologous sequence and deleted most part of the βmaj & βmin gene. 8 strains of positive ES cells cloning were obtained, all of them were testified positive with polymerase chain reaction and Southern blotting. The results of AKP staining, embryoid bodies formation in vitro and teratomas formation in nude mice showed undifferentiated state and pluripotential characteristic of cell line. CONCLUSIONS: A knock-out recombinant plasmic vector of βmaj & βmin of β-globin gene in C129 mouse was constructed, in which a knock-out βmaj & βmin gene of β-globin gene ES-D3 cell line was established by electroporation, characterization of these cells show a prospect utility in producing gene knock-out mice in blastocysts stages. 相似文献
20.
水分胁迫诱导八棱海棠和平邑甜茶细胞程序性死亡的研究 总被引：4，自引：0，他引：4
以苹果属抗旱能力较强的八棱海棠(Malus robusta Rehd. ) 和较弱的平邑甜茶[Malus hupehensis (Pamp.) Rehd. ] 水培幼苗为试验材料, 利用细胞形态学、DNA ladder等技术, 用20% PEG 6000模拟水分胁迫进行诱导细胞程序性死亡研究。经DAP I染色后荧光显微镜观察发现两个苹果属材料均呈现细胞染色质凝集、细胞核缩小、变形、解体等细胞程序性死亡的形态学特征; 同时, DNA琼脂糖凝胶电泳可观察到明显的“梯状”DNA条带(DNA ladder) 。由此表明, 水分胁迫诱导苹果属植物具有明显的细胞程序性死亡的典型形态及生化特征, 是一种主动的细胞死亡过程, 八棱海棠细胞程序性死亡持续的时间长于平邑甜茶。 相似文献