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Sequential extraction of RNA,DNA and protein from cultured cells of the same group
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作者 Ying-Yu Cui 《World Journal of Methodology》 2023年第5期484-491,共8页
BACKGROUND Efficient extraction of nucleic acids and proteins(ENAP)from cells is a prerequisite for precise annotation of gene function,and has become laboratory routine for revealing the mysteries of life.However,cel... BACKGROUND Efficient extraction of nucleic acids and proteins(ENAP)from cells is a prerequisite for precise annotation of gene function,and has become laboratory routine for revealing the mysteries of life.However,cell samples are often from different culture dishes,resulting in inevitable experimental errors and sometimes poor repeatability.AIM To explore a method to improve the efficiency of ENAP,minimizing errors in ENAP processes,enhancing the reliability and repeatability of subsequent experimental results.METHODS A protocol for the sequential isolation of RNA,DNA,and proteins from the same cultured HepG2 cells using RNAzol reagent is presented here.The first step involves culturing HepG2 cells to the exponential phase,followed by the sequential isolation of RNA,DNA,and proteins from the same cultured cells in the second step.The yield of nucleic acids and proteins is detected in the third step,and their purity and integrity are verified in the last step.RESULTS The procedure takes as few as 3-4 d from the start to quality verification and is highly efficient.In contrast to the existing kits and reagents,which are primarily based on independent isolation,this RNAzol reagent-based method is characterized by the sequential isolation of RNA,DNA,and proteins from the same cells,and therefore saves time,and has low cost and high efficiency.CONCLUSION The RNA,DNA,and proteins isolated using this method can be used for reverse transcription-polymerase chain reaction,polymerase chain reaction,and western blotting,respectively. 展开更多
关键词 Sequential extraction Ribonucleic acid Deoxyribonucleic acid PROTEIN cultured cells
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Current status and challenges for cell-cultured milk technology: a systematic review
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作者 Hyuk Cheol Kwon Hyun Su Jung +1 位作者 Vahinika Kothuri Sung Gu Han 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2024年第5期1778-1792,共15页
Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture.While most cellular agriculture is predominantly centered on the produc... Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture.While most cellular agriculture is predominantly centered on the production of cultured meat,there is a growing demand for an understanding of the production techniques involved in dairy products within cellular agriculture.This review focuses on the current status of cellular agriculture in the dairy sector and technical challenges for cell-cultured milk production.Cellular agriculture technology in the dairy sector has been classified into fermentation-based and animal cell culture-based cellular agriculture.Currently,various companies synthesize milk components through precision fermentation technology.Nevertheless,several startup companies are pursuing animal cell-based technology,driven by public concerns regarding genetically modified organisms in precision fermentation technology.Hence,this review offers an up-to-date exploration of animal cell-based cellular agriculture to produce milk components,specifically emphasizing the structural,functional,and productive aspects of mammary epithelial cells,providing new information for industry and academia. 展开更多
关键词 Cell culture system Cell-cultured milk Mammary epithelial cells Precision fermentation
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Transplantation of primary cultured embryonic mesencephalic neural precursor cells for treating Parkinsonian rats 被引量:7
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作者 Li Fei Chengchuan Jiang +2 位作者 Linyin Feng Yaodong Ji Zhongliang Ding 《Neural Regeneration Research》 SCIE CAS CSCD 2006年第1期6-9,共4页
BACKGROUND: Choosing proper donor cells is one of keys in experimental and clinical studies on cell replacement therapy (CRT) for treating Parkinson disease (PD). Embryonic mesencephalic precursor cells (MPCs) ... BACKGROUND: Choosing proper donor cells is one of keys in experimental and clinical studies on cell replacement therapy (CRT) for treating Parkinson disease (PD). Embryonic mesencephalic precursor cells (MPCs) can stably differentiate into dopaminergic neuron after in vitro proliferated culture. As compared with embryonic stem cell and neural stem cell strains, cell composition of embryonic MPCs after primary culture is also the most close to that of embryonic mesencephalic ventral cell suspension without proliferated culture. Successful experience accumulated in the latter suggests that primary cultured embryonic MPCs might be the most potential donor cells in clinical application with CRT for treating PD so far. OBJECTIVE: To investigate the feasibility of primary cultured embryonic precursor cells cultured primarily as donor cells in CRT for treating PD in rats. DESIGN : A randomized and controlled trial taking SD rats as experimental animals.SETTING: Department of Neurosurgery, Huashan Hospital Affiliated to Fudan University.MATERIALS: This experiment was carried out at the Institute of Neuroscience, Shanghai Institute for Biological Science, Chinese Academy of Sciences from July 2003 to June 2004. Totally 26 female SD rats, with body mass of 200 to 220 g, were provided by Shanghai Experimental Animal Center of Chinese Academy of Sciences. METHODS : Stereotaxic injection of 6-hydroxydopamine into the medial forebrain bundle were perfored to develop PD model rat. Among 26 SD rats, 20 rats achieved a more than 5 turns/min in apomorphine induced rotation test, reaching the standard of PD model rats. Immunohistochemical detection was performed on 1 out of 20 model rats after execution, and the other 19 rats were randomly divided into control group (n=5), sham transplantation group (n=5)and cell grafted group (n=9). Primary cultured E12 MPC cell suspension (1.2×10^11 L^-1)were used as donor cells. 4μL primary cultured E12 MPC cell suspension prepared freshly was injected into the lesioned corpus striatum of rats in cell grafted group, and 4μL D-Hank's solution was injected in sham transplantation group in the same way. There was no injection in control group. Apomorphine-induced rotation rate of PD rats were recorded respectively in cell grafted group and sham transplantation group pre-operation (initial value) and at postoperative 2, 4, 6 and 16 weeks. Apomorphine-induced rotation rate of PD rats was recorded in control group at postoperative 2 months (initial value) and following 2,4,6 and 16 weeks. To determine TH antigen with immunohistological ABC method (DAB developing) at 6 months post-transplantation to investigate the differentiation and survival of donor cells in the host body.MAIN OUTCOME MEASURES: Apomorphine-induced rotation behavior before and after transplantation and the survival and differentiation of implanted cells in the host body at 6 months post-transplantation. RESULTS: Among 19 model rats, one rat died after transplantation respectively in the cell grafted group and sham transplantation group; finally 17 model rats entered the stage of result analysis. Relative apomorphine-induced rotation rate was significantly decreased in the cell grafted group as compared with that before transplantation , with significant difference (P 〈 0.01 .P 〈 0.05);the mean value of relative apomorphine-induced rotation rate was significantly decreased at postoperative 16 weeks in cell grafted group as compared with that of corresponding relative rotation rate in control group , also with significant difference (P 〈 0.05).Immunohistological results showed that donor cells could differentiate into large and multi-polar dopaminergic neurons in the host body. CONCLUSION : Primary cultured embryonic MPCs can be used as the donor cells in CRT for treating PD. 展开更多
关键词 CELL FIGURE Transplantation of primary cultured embryonic mesencephalic neural precursor cells for treating Parkinsonian rats
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Expression of Transforming Growth Factor-β in Cultured Normal Human Lens Epithelia Cells 被引量:2
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作者 黄渝侃 魏厚仁 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2004年第3期289-291,共3页
Summary: In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor β (TGF-β), reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemic... Summary: In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor β (TGF-β), reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical methods were used for detection of TGF-β mRNA and protein in cultured normal human LEC. The results showed that a single RT-PCR amplified product about 310bp was obtained, and the sequence was homologous to the known sequence. TGF-β immunostain was positive in the plasma of LEC. It was suggested that normal human LEC could produce TGF-β, and LEC could be affected by TGF-β through autocrine action. 展开更多
关键词 LENS cell culture transforming growth factor reverse transcriptase-polymerase chain reaction IMMUNOHISTOCHEMISTRY
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Isolation of Cultured Endothelial Progenitor Cells in vitro from PBMCs and CD133^+ Enriched Cells 被引量:2
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作者 郑伟红 万亚峰 +4 位作者 马小鹏 李兴睿 杨志芳 殷茜 易继林 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2010年第1期18-24,共7页
Two isolation methods for sorting of endothelial progenitor cells(EPCs):from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive acti... Two isolation methods for sorting of endothelial progenitor cells(EPCs):from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive activities and function in vitro,to provide a reference for clinical application of EPCs.PBMCs from healthy subjects were used either directly for cell culture or for CD133+ sorting.The two groups of cells were cultured in complete medium 199(M199)for 7 to 14 days and the phenotypes of EPCs were an... 展开更多
关键词 endothelial progenitor cells cell culture MACS
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Hexabromocyclododecane-induced Genotoxicity in Cultured Human Breast Cells through DNA Damage 被引量:2
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作者 LI Rui Jing GAO Hui +3 位作者 NA Guang Shui LU Zi Hao YAO Yao YANG Fan 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2017年第4期296-300,共5页
To investigate the genotoxicity and reveal the potential toxicological mechanisms of Hexabromocyclododecane (HBCD), human breast cells HBL-100 were exposed to a sequence of HBCD concentrations (0, 5, 10, and 50 mg/... To investigate the genotoxicity and reveal the potential toxicological mechanisms of Hexabromocyclododecane (HBCD), human breast cells HBL-100 were exposed to a sequence of HBCD concentrations (0, 5, 10, and 50 mg/L) for 24 h. With a series of zymology and molecular biology methods, we found that HBCD induced dose-dependent oxidative stress on HBL-100 DNA. As revealed in q RT-PCR, activated prognostic factor ATM down-regulated tumor suppressor gene BRCA1 and prompted DNA repair genes h OGG1 and h MTH1 expression in lower concentrations of HBCD (〈 10 mg/L). However, DNA repair were inhibited as well as cell proliferation rate by higher concentrations of HBCD (50 mg/L). The results inferred that the genotoxicity of HBCD was dose-dependent and related to DNA repair pathway. 展开更多
关键词 DNA HBCD Hexabromocyclododecane-induced Genotoxicity in cultured Human Breast cells through DNA Damage
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A study of gentamicin injury mechanisms using cultured mouse cochlear spiral ganglion cells 被引量:1
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作者 GU Xi LIN Chang ZHANG Rong 《Journal of Otology》 2011年第1期31-35,共5页
Objective To study gentamicin injury mechanisms using postnatal mouse cochlear spiral gangcells (SGC). Methods SGCs were isolated using a combinatorial approach of enzymatic digestion and mechanical separation from ... Objective To study gentamicin injury mechanisms using postnatal mouse cochlear spiral gangcells (SGC). Methods SGCs were isolated using a combinatorial approach of enzymatic digestion and mechanical separation from P2 - 6 Kunming mouse cochleae. After 4 days, cultured SGCs were fixed with 4% paraformaldehyde at room temperature for immunocytochemical examination using the methods of S-P and the monoclonal antibody against mouse neurofilament protein (Neurofilament-68/200Kda, NF-L+ H). SGCs were randomly divided into a blank control group and three gentamicin treatment groups (medium gentamicin concentration at 50 mg/L, 100 mg/L and 150 mg/L respectively), SGCs were collected and examined under a transmission electron microscope after being cultured for 48 h. Results SGC primary culture was successful. SGC cytoplasm and neurites were dyed brownish yellow by the monoelonal mouse neurofilament protein antibody. SGCs showed classical bipolar neuron appearance. Under the transmission electron microscope,.gentamicin treated SGCs showed morphological features different compared to those in the blank control group, which might indicate apoptosis. Conclusion Our results indicate that gentamicin has direct toxic effects on cochlear SGCs in mice and the injury mechanism is closely related with apoptosis. Damage to mitochor, dria may play an important role in the process. 展开更多
关键词 GENTAMICINS spiral ganglion cells cultured MICROSCOPY electron transmission apoptosis
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Manganese enhances the expression of the manganese superoxide dismutase in cultured primary chick embryonic myocardial cells 被引量:3
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作者 QIN Shi-zhen LIAO Xiu-dong +4 位作者 LU Lin ZHANG Li-yang XI Lin GUO Yan-li LUO Xu-gang 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第9期2038-2046,共9页
In the present study, the effect of manganese(Mn) on antioxidant status and the expression of the manganese superoxide dismutase(MnSOD) gene in cultured primary myocardial cells collected from the chick embryos wa... In the present study, the effect of manganese(Mn) on antioxidant status and the expression of the manganese superoxide dismutase(MnSOD) gene in cultured primary myocardial cells collected from the chick embryos was investigated. The hypothesis that Mn supplementation would enhance the expression of MnSOD in cultured primary myocardial cells of chick embryos was tested. Eggs collected from Mn-depleted Arbor Acres laying breeder hens were incubated for 10 days and then myocardial cells were isolated and cultivated for 8 days. The embryonic myocardial cells on day 6 were treated with Mn in the cell culture medium at different time points when the proportion of cells showing spontaneous contraction was over 95% after the 3-day primary culture. A completely randomized design involving a 3 Mn levels(0, 0.5 and 1.0 mmol L^(-1))×3 incubation time points(12, 24 and 48 h) factorial arrangement of treatments(n=6) was used in the current experiment. The results showed that MnSOD activity and m RNA expression level were induced by Mn and increased with incubation time, which supported the hypothesis that Mn would enhance the expression of the MnSOD gene, and thus might protect myocardial cells from oxidative stress during the chick embryonic development. 展开更多
关键词 manganese MnSOD expressions cultured primary myocardial cells chick embryos
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THE CHARACTERISTICS OF ENDOGENOUS OUABAIN SECRETION FROM CULTURED BOVINE ADRENOCORTICAL CELLS 被引量:2
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作者 曲毅 吕卓人 《Academic Journal of Xi'an Jiaotong University》 2000年第1期31-33,49,共4页
Objective To compare the characteristics or endogenous ouabaln(EO) secretion with the other adrenocortical hormones and determine the effects of angiotensin I (Ang I ), and ad reno corticotrophin (ACTH ) on the secret... Objective To compare the characteristics or endogenous ouabaln(EO) secretion with the other adrenocortical hormones and determine the effects of angiotensin I (Ang I ), and ad reno corticotrophin (ACTH ) on the secretion or EO. Methods EO was measured by radioimmunoassay from primary cultured bovine adrenocotical cells (BAC). Results oouabain was determined in the media or cultured BAC. Both EO and aldosterone secretion were decreased from the 6uter to inner layer or the cultured adrenal cortex, and the responses to Ang I and ACTH were hlgher than that in the mld layer (P <o. o5) and inner layer (P <o. o1 ). Cortisol secretion was activated by Ang I or ACTH was slgnificantly higher in the mid layer and in the inner layer than that in the outer layer. The tlme-course experlment showed that the gradually rising amounts or aldosterone and cortisol could be determined during the continuous incubation to 48h wlth or without Ang I or ACTH. However, EO did not increase continuously arter 24h or incubation in the basal secreting sltuation and arter 12h of lncubatiou in the stimulating situation by Ang,or ACTH. There were obvlous drops in aldosterone and cortisol secretlou from 3rd day during a 21 day-perlod cell culture, but the peak secretion of ouabain was in 7th day. Conclusion it suggests that the secretory mechanism might be different between EO and aldosterone or cortisol. Also, Ang I and ACTR might be involved in the regulation of Eo secretion. 展开更多
关键词 endogenous ouabain adrenocortical hormonel cell culture
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The Biological Study of the Cultured Human Lens Epithelial Cells in Vitro 被引量:1
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作者 Haike Guo,Shaozhen Li, Xinyuan Cao, Shunwei LiangDepartment of Ophthalmology, The Second Teaching Hospital, Guangzhou Medical College Guangzhou 510260, ChinaZhongshan Ophthalmic Center , Sun Yet-Sen University of Medical Sciences Guangzhou 510060, China 《眼科学报》 1994年第1期27-31,共5页
The human lens epithelial cells (HLE) cultured in vitro was established in normal and cataractous lenses. The biological feature, histological characteristics and the ultrastructure of the cultured HLE cells were inve... The human lens epithelial cells (HLE) cultured in vitro was established in normal and cataractous lenses. The biological feature, histological characteristics and the ultrastructure of the cultured HLE cells were investigated. The results reveal that the proliferative capacity of the culutured HLE cells is reversely proportional to the donour age; the cultured HLE cells has the limited proliferative capacity in vitro. The relieve of the contact inhibition is the effective trigger of the HLE cell prolife... 展开更多
关键词 cell culture LENS CATARACT
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Comparison of biological behavior of lacrimal gland adenoid cystic carcinoma with high-grade transformation cells 被引量:2
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作者 Chuan-Li Zhang Li-Min Zhu +3 位作者 Xun Liu Mei-Xia Jiang Ting-Ting Lin Yan-Jin He 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2023年第2期163-171,共9页
AIM:To evaluate the differences between human lacrimal gland adenoid cystic carcinoma with high-grade transformation(LACC-HGT)primar y cells cultured by high-grade transformation tissue and non-high-grade transformati... AIM:To evaluate the differences between human lacrimal gland adenoid cystic carcinoma with high-grade transformation(LACC-HGT)primar y cells cultured by high-grade transformation tissue and non-high-grade transformation(non-HGT)primary cells cultured by non-highgrade transformation tissue in proliferation,metastasis,drug susceptibility,and genes.METHODS:LACC-HGT primary cells were established by tissue block culture,and the 4^(th)to 10^(th)generation primary cells were selected as research objects.The cells were preliminarily identified by immunofluorescent staining.The differences between non-HGT and LACC-HGT primary cells in terms of proliferation,metastasis,and drug susceptibility were compared by cell counting kit-8(CCK-8)assay,wound healing,and drug sensitivity experiments.Differentially expressed genes were screened using mRNA array.Gene expression was analyzed using real-time quantitative polymerase chain reaction(RT-qPCR).RESULTS:LACC-HGT primary cells were successfully cultured by tissue block culture.Immunofluorescence staining results showed that cytokeratin(CK)and CK7 expression levels were positive in LACC-HGT primary cells.CCK-8 results showed that the proliferation ability of LACCHGT cells was significantly higher than that of non-HGT cells.Wound healing experiment showed that the migration ability of LACC-HGT cells was significantly higher than that of non-HGT cells.LACC-HGT cells were also less sensitive to cisplatin and paclitaxel than non-HGT cells.Compared with non-HGT cells,9566 differentially expressed genes were found in LACC-HGT primary cells,of which 5162 were upregulated and 4404 were down-regulated.The expression of N-acetylneuraminate pyruvate lyase(NPL),MARVEL domain containing 3(MARVELD3),syntabulin(SYBU),and allograft inflammatory factor 1(AIF1)was higher in LACCHGT cells than in non-HGT cells,whereas that of periostin(POSTN)was lower.CONCLUSION:LACC-HGT primary cells have faster proliferation,stronger migration ability,and poorer sensitivity to chemotherapy drugs than non-HGT primary cells.The expression of mRNAs in non-HGT and LACC-HGT primary cells are significantly different.These features are speculated to be the reasons why high-grade transformation tissues exhibit higher malignant degree and poorer prognosis than their counterparts. 展开更多
关键词 lacrimal gland lacrimal gland adenoid cystic carcinoma high-grade transformation primary cell culture biological behavior mRNA array
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Antagonistic Effects of Tranilast on Proliferation and Collagen Synthesis Induced by TGF-β_2 in Cultured Human Trabecular Meshwork Cells
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作者 笪邦红 曹阳 +3 位作者 魏厚仁 陈志新 水迎波 李忠玉 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2004年第5期490-492,496,共4页
Whether tranilast had antagonistic effect on proliferation inhibition and collagen synthesis promotion induced by TGF-β 2 in cultured human trabecular meshwork cells was investigated. Suspension of 1×104 cultur... Whether tranilast had antagonistic effect on proliferation inhibition and collagen synthesis promotion induced by TGF-β 2 in cultured human trabecular meshwork cells was investigated. Suspension of 1×104 cultured human trabecular meshwork cells of 3—5 passage was distributed in each well of a 96-well disk and divided into control group and experimental group. After 24 h, 0 μg/ml (control), 12.5 μg/ml, 25 μg/ml, 50 μg/ml tranilast with 3.2 ng/ml TGF-β 2 were added into the incubation medium. Another 24 h later, proliferation and collagen synthesis in cultured human trabecular meshwork cells were examined respectively by using tetrazolium-based semiautomated colormetric (MTT) assay and 3H-proline incorporation with liquid scintillation technique. The results showed absorbance (A) values of the experimental groups were 0.9036±0.3017, 1.1361± 0.1352, 1.2457±0.1524 according to the different concentrations of tranilast, and 0.8956± 0.1903 of the control group. In comparison with the control group, 25 μg/ml (q'=3.23, P< 0.05), 50 μg/ml (q'=4.70, P<0.01) tranilast significantly antagonized the decrease of the A values induced by TGF-β 2 in the cultured human trabecular meshwork cells. In comparison with the control group [817.37±124.21 cpm/104 cells], 12.5 μg/ml (620.33±80.46 cpm/104 cells, q'= 4.26, P< 0.05), 25 μg/ml (594.58±88.13 cpm/104 cells, q'=4.81, P<0.01), 50 μg/ml (418.64±67.90 cpm/104 cells, q'=8.62, P<0.01) tranilast significantly inhibited the incorporation of 3H-proline into the cultured human trabecular meshwork cells promoted by TGF-β 2 in a dose-dependent manner. It was concluded that tranilast had the antagonistic effect on the proliferation inhibition and collagen synthesis promotion induced by TGF-β 2 in the cultured human trabecular meshwork cells. 展开更多
关键词 transforming growth factor HUMAN trabecular meshwork cultured cell PROLIFERATION COLLAGEN
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Effect of Transforming Growth Factor-β_2 on Phagocytosis in Cultured Bovine Trabecular Meshwork Cells
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作者 曹阳 魏厚仁 +1 位作者 笪邦红 黄毅 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2001年第4期318-320,共3页
The effect of transforming growth factor β 2 (TGF β 2) on phagocytosis in bovine trabecular meshwork cells in vitro was investigated. After the cultured bovine trabecular meshwork cells were treated with 0 ... The effect of transforming growth factor β 2 (TGF β 2) on phagocytosis in bovine trabecular meshwork cells in vitro was investigated. After the cultured bovine trabecular meshwork cells were treated with 0 ng/ml, 0.32 ng/ml, 1 ng/ml, 3.2 ng/ml TGF β 2 for 24 h, latex beads were added into the incubation medium, and the numbers of the latex beads in 20 adjacent cells were counted under a microscope 24 h later, after treatment with Wright's stain. Our results showed that the average numbers of the latex beads in the trabecular meshwork cells treated with TGF β 2 of different concentrations were 53.1±1.7 beads/cell, 56.4±2.9 beads/cell and 77.9±6.5 beads/cell respectinvely, in comparison with 45.5±3.3 beads/cell of the control group. TGF β 2 significantly increased the number of the latex beads phagocytosed by cultured bovine trabecular meshwork cells in a dose dependent manner. TGF β 2 could promote the phagocytosis of bovine trabecular meshwork cells in vitro . It may be involved in the cellularity decrease of the trabecular meshwork in the patients of primary open angle glaucoma through promoting the phagocytosis of trabecular meshwork cells. 展开更多
关键词 transforming growth factor trabecular meshwork cultured cells PHAGOCYTOSIS
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The Effect of Zinc on the Apoptosis of Cultured Human Retinal Pigment Epithelial Cells
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作者 陈雯 王智 张缨 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2003年第4期414-417,共4页
To clarify the effects of zinc on the proliferation and apoptosis of cultured human retinal pigment epithelia (RPE) and the expression of caspase 3 in RPE cells. The effect of Zinc on theproliferation of RPE were exa... To clarify the effects of zinc on the proliferation and apoptosis of cultured human retinal pigment epithelia (RPE) and the expression of caspase 3 in RPE cells. The effect of Zinc on theproliferation of RPE were examined with MTT method. TUNEL method was used to detect the apoptosis of RPE cells. Caspase 3 was detected by immunohistochemistry. A concentration of zinc higher than 0.001 μM could inhibit the proliferation of RPE. And the relationship between concentration of zinc higher than 10 μM and growth prohibition rate of RPE cells was dose dependent. All concentrations of zinc including 0.001 μM enhanced the expression of caspase 3 of RPE. But only the concentration of zinc higher than 0.01 μM could induce apoptosis of RPE. It is concluded that zinc could enhance the expression of caspase 3 of RPE cells and induce apoptosis of RPE cells. Caution should be taken when using zinc supplements for the treatment of ARMD patients without deficiency of zinc. 展开更多
关键词 PIGMENT EPITHELIUM EYES cell culture APOPTOSIS ZINC
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Alcohol Dehydrogenase Activity in Cultured Cells from Different Tobacco (Nicotiana tabacum L.) Varieties
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作者 ZHENG Ling, YANG Yue sheng ,ZHENG Ying dong College of Biotechnology, South China Agricultural University, Guangzhou 510642,China 《Wuhan University Journal of Natural Sciences》 CAS 1999年第4期495-497,共3页
The activity of alcohol dehydrogenase (ADH) in cultured cells of various tobacco was determined. It was found that significant differences existed in cells of different varieties cultured under normal conditions and a... The activity of alcohol dehydrogenase (ADH) in cultured cells of various tobacco was determined. It was found that significant differences existed in cells of different varieties cultured under normal conditions and as well after treated with exogenous ethanol. The ADH activity had positive relation with the ability of the cells to catabolize exogenous ethanol, indicating that the main function of the ADH in tobacco cells was in the direction of converting ethanol to acetaldehyde. 展开更多
关键词 alcohol dehydrogenase suspension cultured cells TOBACCO
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T-type calcium channel expression in cultured human neuroblastoma cells
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作者 Xianjie Wen Shiyuan Xu +4 位作者 Lingling Wang Hua Liang Chengxiang Yang Hanbing Wang Hongzhen Liu 《Neural Regeneration Research》 SCIE CAS CSCD 2011年第31期2410-2413,共4页
Human neuroblastoma cells (SH-SY5Y) have similar structures and functions as neural cells and have been frequently used for cell culture studies of neural cell functions.Previous studies have revealed L-and N-type c... Human neuroblastoma cells (SH-SY5Y) have similar structures and functions as neural cells and have been frequently used for cell culture studies of neural cell functions.Previous studies have revealed L-and N-type calcium channels in SH-SY5Y cells.However,the distribution of the low-voltage activated calcium channel (namely called T-type calcium channel,including Cav3.1,Cav3.2,and Cav3.3) in SH-SY5Y cells remains poorly understood.The present study detected mRNA and protein expres-sion of the T-type calcium channel (Cav3.1,Cav3.2,and Cav3.3) in cultured SH-SY5Y cells using real-time polymerase chain reaction (PCR) and western blot analysis.Results revealed mRNA and protein expression from all three T-type calcium channel subtypes in SH-SY5Y cells.Moreover,Cav3.1 was the predominant T-type calcium channel subtype in SH-SY5Y cells. 展开更多
关键词 calcium ion channel calcium signaling cell culture neural regeneration real-time PCR T-type calcium channel western blot neural regeneration
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STUDY ON DIFFERENTIATION OF RATS EMBRYONIC STEM CELLS CULTURED IN BRL-CM INTO NEURAL PRECURSOR CELLS
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作者 张晓智 李旭 +1 位作者 徐海伟 陈葳 《Journal of Pharmaceutical Analysis》 SCIE CAS 2003年第1期55-58,共4页
Objective To investigate whether buffalo rat liver cell conditioned medium (BRL CM) can be used as the culture medium of embryonic stem (ES) cells, and to get relatively pure neural precursor cells (NPCs) for treat... Objective To investigate whether buffalo rat liver cell conditioned medium (BRL CM) can be used as the culture medium of embryonic stem (ES) cells, and to get relatively pure neural precursor cells (NPCs) for treatment aim. Methods Mouse ES cells were cultured in BRL CM and medium contain leukemia inhibitory factor (LIF), respectively. NPCs were selectively cultured in serum free medium. Alkaline phosphatase activity was visualized with NBT/BCIP and nestin antigen was detected with immunocytochemical methods. Results BRL CM could be used as an efficiency culture condition instead of LIF in ES cells culture. About 86% of cells derived from ES cells in the serum free culture were NPCs. Conclusion BRL CM can replace LIF to use in ES cell culture. High purity of NPC can be induced from ES cells with serum free culture method. 展开更多
关键词 embryonic stem cell neural precursor cell cell culture
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A Primary Study of the Proliferation Sti mulation Effect by Pyrroloquinoline Quinone (PQQ) on Cultured Schwann Cells
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作者 Hao-Huan LI~1 Shi-Qing LIU~1 Jing-Ping OU YANG~2 Hao PENG~1Yi XU~2 Hai-Lu YANG~2 Fei ZENG~3 1(Orthopedic Department of Renmin Hospital,Wuhan University, Wuhan 430060, China)2(Pathophysiology Department, School of Medicine, Wuhan University, Wuhan 430071, China)3(Neurology Department of Renmin Hospital of Wuhan University, Wuhan 430060, China) 《生物医学工程学杂志》 EI CAS CSCD 北大核心 2005年第S1期79-80,102,共3页
关键词 PQQ A Primary Study of the Proliferation Sti mulation Effect by Pyrroloquinoline Quinone on cultured Schwann cells
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Expression of Inositol 1,4,5-trisphosphate Receptor mRNA in Myocardium of Spontaneous Hypertension Rats and Cultured Vascular Smooth Muscle Cells of Rats
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作者 刘乃丰 张寄南 +3 位作者 耿茜 杨笛 董莉 马文珠 《Journal of Nanjing Medical University》 2002年第2期75-79,共5页
Objective\ To investigate expression of inositol 1,4,5 trisphosphate receptor (IP\-3R) mRNA on sacroplasmic reticular in myocardium of spontaneous hypertension rats (SHRs) and cultured vascular smooth muscle cells (V... Objective\ To investigate expression of inositol 1,4,5 trisphosphate receptor (IP\-3R) mRNA on sacroplasmic reticular in myocardium of spontaneous hypertension rats (SHRs) and cultured vascular smooth muscle cells (VSMC) of rats and effects of perindopril and urapidil on them. Methods\ SHRs were orally given perindopril (1.0 mg·kg\+\{ 1\}·d\+\{ 1\}) or urapidil (15 mg·kg\+\{ 1\}·d\+\{ 1\}) for 24 weeks, respectively. Expression of IP\-3R mRNA was examined by semi quantitative reverse transcription polymers chain reaction (RT PCR) using three oligonuclotide primers for each subtype of IP\-3R with β actin as internal label. Results\ All subtypes of IP\-3R were expressed in myocardium of SHR, WKY and cultured VSMC. Expression of IP\-3R mRNA in left ventricle of SHR was markedly enhanced. Urapidil could down regulate expression of IP\-3R Ⅰand IP\-3R Ⅲ, perindopril slightly increased expression of IP\-3R Ⅱ and decreased expression of IP\-3R Ⅰand IP\-3R Ⅲ in myocardium of SHR. Conclusion\ Our results suggest that expression of IP\-3R mRNA in cardiovascular system could be regulated by urapidil and perindopril. 展开更多
关键词 calcium release channel signal transduction inositol 1 4 5 trisphosphate receptor spontaneous hypertension rat vascular smooth muscle cultured cells polymers chain reaction
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Effect of captopril or verapamil on intracellular sodium in cultured vascular smooth muscle cells
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作者 QI JIAN HUA LU ZHANG +3 位作者 JUN WANG MIN LU XIN MING WANG ZHENG JUN JIN(Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.)(Department of Pharmacology, Shanghai Second Medical University, Shanghai 200025, China.) 《Cell Research》 SCIE CAS CSCD 1996年第1期47-53,共7页
The effects of captopril (Cap) and verapamil (Ver)alone and in combination on intracellular Na+ concentration ([Na+]i) in cultured aortic smooth muscle cells (ASMC) of rabbits was evaluated by a direct measurement of ... The effects of captopril (Cap) and verapamil (Ver)alone and in combination on intracellular Na+ concentration ([Na+]i) in cultured aortic smooth muscle cells (ASMC) of rabbits was evaluated by a direct measurement of [Na+]i with fluorescent dye sodium-binding benzofuran isophthalate (SBFI) combined with digital image. [Na+]i in resting cells was found to be 11.9 ± 0. 7 mmol/L. Angiotensin II (Ang-II, 0.1-10μmol/L) induced an increase of [Na+]i in concentration-dependent manner. Ver (0.1-10μmol/L) inhibited Ang-II (1 μmol/L)-induced increase in [Na+]i, while Cap enhanced Ang-II-induced increase in [Na+]i at 10μmol/L but not at 0.1-1μmol/L. Ver (0.1-1μmol/L)abolished enhancement of Ang-II-induced increase in [Na+]i by Cap. Thus, the inhibition of Capenhanced [Na+]i by Ver may provide a new hypothesis for the underlying molecular mechanism of synergistic effect of the combination of Ca2+ antagonists and angiotensinconverting enzyme inhibitors in controlling blood pressure. 展开更多
关键词 cultured aortic smooth muscle cells angiotensin CAPTOPRIL VERAPAMIL sodiumbinding benzofuran isophthalate
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