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Postnatal development of rat retina:a continuous observation and comparison between the organotypic retinal explant model and in vivo development
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作者 Baoqi Hu Rui Wang +8 位作者 Hanyue Zhang Xiou Wang Sijia Zhou Bo Ma Yan Luan Xin Wang Xinlin Chen Zhichao Zhang Qianyan Kang 《Neural Regeneration Research》 SCIE CAS 2025年第3期900-912,共13页
The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and contin... The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation. 展开更多
关键词 bipolar cells differentiation in vivo microglia Müller glia organotypic retinal explant culture postnatal retina development proliferation retinal progenitor cells
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Punch‑excised explants of bovine mammary gland to model early immune response to infection
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作者 Pablo Gomes Noleto Florence B.Gilbert +4 位作者 Christelle Rossignol Patricia Cunha Pierre Germon Pascal Rainard Rodrigo Prado Martins 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2023年第6期2649-2662,共14页
Background Mammary gland(MG)infections(mastitis)are frequent diseases of dairy cows that affect milk quality,animal welfare and farming profitability.These infections are commonly associated with the bacteria Escheric... Background Mammary gland(MG)infections(mastitis)are frequent diseases of dairy cows that affect milk quality,animal welfare and farming profitability.These infections are commonly associated with the bacteria Escherichia coli and Staphylococcus aureus.Different in vitro models have been used to investigate the early response of the MG to bacteria,but the role of the teat in mastitis pathogenesis has received less attention.In this study,we used punch-excised teat tissue as an ex vivo model to study the immune mechanisms that arise early during infection when bacteria have entered the MG.Results Cytotoxicity and microscopic analyses showed that bovine teat sinus explants have their morphology and viability preserved after 24 h of culture and respond to ex vivo stimulation with TLR-agonists and bacteria.LPS and E.coli trigger stronger inflammatory response in teat when compared to LTA and S.aureus,leading to a higher produc-tion of IL-6 and IL-8,as well as to an up-regulation of proinflammatory genes.We also demonstrated that our ex vivo model can be applied to frozen-stored explants.Conclusions In compliance with the 3Rs principle(replacement,reduction and refinement)in animal experimenta-tion,ex vivo explant analyses proved to be a simple and affordable approach to study MG immune response to infec-tion.This model,which better reproduces organ complexity than epithelial cell cultures or tissue slices,lends itself particularly well to studying the early phases of the MG immune response to infection. 展开更多
关键词 Bovine teat Escherichia coli explant MASTITIS Staphylococcus aureus 3Rs
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辣椒离体培养及再生体系的建立 被引量:2
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作者 李浩宇 贾利 +7 位作者 宋婷婷 汪胜 张于 方凌 张其安 严从生 隋益虎 江海坤 《中国蔬菜》 北大核心 2024年第1期38-45,共8页
为了建立高效的辣椒离体再生体系,选择5种不同基因型的辣椒无菌苗,研究了添加不同激素配比的MS培养基对芽诱导分化、芽伸长增殖、生根和移栽等的影响。结果表明:MS+6.0 mol·L^(-1)6-BA+1.0 mol·L^(-1)IAA+5.0 mol·L^(-1)... 为了建立高效的辣椒离体再生体系,选择5种不同基因型的辣椒无菌苗,研究了添加不同激素配比的MS培养基对芽诱导分化、芽伸长增殖、生根和移栽等的影响。结果表明:MS+6.0 mol·L^(-1)6-BA+1.0 mol·L^(-1)IAA+5.0 mol·L^(-1)Ag NO_(3)组合为芽分化的最适培养基,可诱导出白色松软的愈伤组织,Flamingo-bill诱导率为100.0%,子叶诱导率为86.7%。MS+3.0mol·L^(-1)6-BA+2.0 mol·L^(-1)IAA+5.0 mol·L^(-1)Ag NO3+1.5 mol·L^(-1)GA3组合为不定芽伸长的最适培养基,Flamingo-bill不定芽伸长率为66.7%,子叶不定芽伸长率为30.0%。MS+0.5 mol·L^(-1)NAA组合为不定芽生根的最适培养基,Flamingo-bill和子叶不定芽生根率均为100.0%。通过Flamingo-bill和子叶外植体均能获得再生植株。 展开更多
关键词 辣椒 离体培养 植株再生 外植体
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元宝枫组织培养与快速繁殖技术 被引量:1
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作者 权志珣 燕丽萍 +4 位作者 王因花 仲伟国 梁燕 吴德军 马秋月 《中南林业科技大学学报》 CAS CSCD 北大核心 2024年第1期119-127,139,共10页
【目的】元宝枫是我国特有的木本油料植物,非常适合开发利用,但其组培离体繁殖生根非常困难,建立合理的完整组培离体再生体系,在短时期内获得大量种苗和当年生带芽茎段,为该树种优良种苗规模化生产提供新途径,为选育元宝枫良种、种质资... 【目的】元宝枫是我国特有的木本油料植物,非常适合开发利用,但其组培离体繁殖生根非常困难,建立合理的完整组培离体再生体系,在短时期内获得大量种苗和当年生带芽茎段,为该树种优良种苗规模化生产提供新途径,为选育元宝枫良种、种质资源保存与推广等提供研究基础。【方法】以元宝枫当年生带芽茎段和种子为试材,通过正交设计筛选法和单因素筛选法对其灭菌方式及腋芽诱导、继代增殖、生根培养基进行设计筛选。在MS、1/2MS和NN69基础培养基中分别添加不同质量浓度的6-BA、NAA和TDZ激素,通过不同配比找出元宝枫各阶段的最适培养基。【结果】全年中4月份为带芽茎段最佳采集时间,经洗衣粉溶液轻刷表面并冲洗半小时后,以75%酒精30 s+0.1%HgCl_(2) 10 min浸泡处理的灭菌效果最佳。再生体系各阶段最适培养基均为MS基础培养基,在其基础上再添加不同质量浓度的激素可以诱导嫩芽分化成不同器官。启动培养阶段,6-BA对腋芽诱导影响最大,最适培养基诱导率可达73.33%;继代增殖阶段,TDZ对不定芽分化影响最大,增殖系数最高可达4.31;生根阶段,采用单因素筛选法,发现添加0.2 mg·L^(-1) IBA元宝枫生根率和生根条数均为最高,达93.33%和4.3。【结论】元宝枫最适培养基为:1)启动培养:MS+0.06 mg·L^(-1) 6-BA和0.06 mg·L^(-1) NAA,诱导率为73.33%;2)增殖培养:MS+0.30 mg·L^(-1) TDZ和0.05 mg·L^(-1) NAA;3)生根培养:MS+0.2 mg·L^(-1) IBA。 展开更多
关键词 元宝枫 外植体 组织培养 快速繁殖
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利用绿色荧光蛋白优化辣椒遗传转化体系
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作者 董依萍 刘画 +3 位作者 刘丹 周迎佳 李峰 邓颖天 《广东农业科学》 CAS 2024年第3期103-113,共11页
【目的】农杆菌介导的辣椒遗传转化较为困难,至今仍未建立高效转化体系。绿色荧光蛋白(GFP)基因是植物遗传转化中常用的报告基因,以GFP为报告基因,优化农杆菌介导的辣椒遗传转化体系。【方法】利用GFP表达系统,统计3个辣椒品种(‘HP’‘... 【目的】农杆菌介导的辣椒遗传转化较为困难,至今仍未建立高效转化体系。绿色荧光蛋白(GFP)基因是植物遗传转化中常用的报告基因,以GFP为报告基因,优化农杆菌介导的辣椒遗传转化体系。【方法】利用GFP表达系统,统计3个辣椒品种(‘HP’‘8214’和‘L55’)中3种不同外植体(子叶、下胚轴和Flamingo-bill外植体)的不定芽分化率、不定根分化率与荧光阳性率,探究农杆菌侵染浓度、侵染时间、预培养时间和共培养时间等因素对不定芽、不定根分化率及荧光阳性率的影响。【结果】3个辣椒品种的Flamingo-bill外植体不定芽分化率均显著高于下胚轴与子叶外植体,其中‘L55’Flamingo-bill外植体的不定芽分化率最高、达77.59%,故选用‘L55’Flamingo-bill外植体进行后续研究。4种农杆菌不同侵染浓度和时间组合下,‘L55’Flamingo-bill外植体均可产生不定芽、不定根及表达GFP的愈伤组织,当农杆菌侵染浓度为OD_(600)=0.05,侵染时间为30 min时,不定芽分化率和荧光阳性率最高,分别达48.39%、4.84%。在6种不同预培养与共培养时间组合处理下,Flamingo-bill外植体也均产生不定芽和不定根,预培养1 d、共培养1~2 d处理下的不定芽分化率和荧光阳性率最高,分别达48.44%、12.50%。最后对表达GFP荧光的不定根与愈伤组织进行PCR检测,结果显示GFP、Kan和Cas9基因在荧光阳性的组织中均被检测到,表明农杆菌介导的T-DNA插入成功且转化稳定。【结论】辣椒的外植体类型、农杆菌侵染浓度、侵染时间、预培养和共培养时间均对辣椒遗传转化效率有影响。以GFP为报告基因,筛选合适的转化条件可提高农杆菌介导的辣椒遗传转化效率。 展开更多
关键词 辣椒 flamingo-bill外植体 绿色荧光蛋白 遗传转化 农杆菌 组织培养
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君迁子组培体系的建立及优化
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作者 吕中一 闻家乐 +5 位作者 刘泽远 张馨予 胡碧春 范芝蕊 关长飞 杨勇 《西北农林科技大学学报(自然科学版)》 CSCD 北大核心 2024年第5期102-109,共8页
【目的】建立君迁子(Diospyros lotus L.)组培快繁体系并进行优化,为君迁子组培苗的生产应用提供技术支持及理论依据。【方法】以污染率、褐化率、成活率为考察指标,设置体积分数10%H 2O 2消毒10,20,30 min和体积分数2%NaClO消毒5 min+... 【目的】建立君迁子(Diospyros lotus L.)组培快繁体系并进行优化,为君迁子组培苗的生产应用提供技术支持及理论依据。【方法】以污染率、褐化率、成活率为考察指标,设置体积分数10%H 2O 2消毒10,20,30 min和体积分数2%NaClO消毒5 min+体积分数10%H 2O 2消毒10 min 4个处理,筛选君迁子外植体适宜的消毒方法;分别以君迁子萌动芽、春梢、腋芽和休眠芽作为外植体,筛选最合适的外植体材料;分析DKW、(1/2N)MS、1/2MS、MS培养基对组培苗启动培养的影响,筛选组培苗生长的最适培养基;增殖培养中,激素ZT、6-BA均设0,1.0,2.0,3.0,4.0 mg/L 5个水平,IAA设0.05,0.1,0.2 mg/L 3个水平,筛选组培苗继代增殖最适激素配比;最后分析不同质量浓度IBA(0,1.0,2.0,3.0 mg/L)对组培苗生根的影响。【结果】君迁子外植体最适的消毒方法为体积分数2%NaClO消毒5 min+体积分数10%H 2O 2消毒10 min;最佳外植体材料为萌动芽,其成活率可达60.00%;在(1/2N)MS培养基上组培苗生长更加健壮,启动培养效果较好;增殖培养最佳的激素配比为3.0 mg/L ZT+0.1 mg/L IAA,在此处理下,组培苗2级愈伤占比79.00%,显著高于其余处理,2级和3级株高占比分别为55.33%和44.67%,长势均匀且主茎粗壮,其增殖系数达5.83;君迁子生根培养较为适宜的IBA质量浓度为1.0 mg/L,生根率可达83.33%。【结论】初步建立了较为完整的君迁子组培体系,有助于柿砧木无性系产业化扩繁的推进。 展开更多
关键词 君迁子 外植体 组织培养 褐化 生根
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濒危植物粗梗水蕨孢子高效无菌萌发及成苗技术
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作者 杨星宇 梁玉婷 +3 位作者 陈永勤 田姗姗 陈益娴 钟昌龙 《湖北大学学报(自然科学版)》 CAS 2024年第4期584-590,共7页
粗梗水蕨(Ceratopteris pteridoides)为一年生蕨类植物,是水体和湿地生态系统中重要成员之一。上世纪80年代以来,由于生存环境受到了严重破坏,我国粗梗水蕨的数量锐减。虽然早在1999年粗梗水蕨就被国务院列为国家二级重点保护濒危植物,... 粗梗水蕨(Ceratopteris pteridoides)为一年生蕨类植物,是水体和湿地生态系统中重要成员之一。上世纪80年代以来,由于生存环境受到了严重破坏,我国粗梗水蕨的数量锐减。虽然早在1999年粗梗水蕨就被国务院列为国家二级重点保护濒危植物,但自然界中粗梗水蕨还是越来越稀少,对其进行人工保护势在必行。我们研究并解决了粗梗水蕨孢子无菌培养中的一些主要问题,建立了一套简单、高效的孢子萌发成苗方法。首先,用0.5%有效氯的次氯酸钠消毒孢子5 min,在含3 mL/L抗菌剂Plant Preservation Mixture^(TM)(PPM)和20 g/L蔗糖的1/2 MS液体培养基中培养5周;然后,将孢子萌发形成的培养物转到含20 g/L糖和6 g/L琼脂的1/2 MS培养基上培养1个月,得到大量的幼孢子体(幼苗);最后,将幼苗转到新鲜的含20 g/L糖和6 g/L琼脂的1/2 MS培养基上,1个月后可长成健壮的大苗。2021年组培苗投放到野外保护区后能正常生长和形成孢子,次年部分孢子自然萌发形成了新的植株。 展开更多
关键词 粗梗水蕨 濒危植物保护 外植体消毒 孢子无菌萌发
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悬铃木组织培养再生体系的建立
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作者 孟汇然 郭尚敬 +3 位作者 张演义 隋超 王德颖 吕陆佳 《现代农业科技》 2024年第8期78-80,共3页
为了优化建立悬铃木组织培养再生体系,本研究以健壮、无病虫害的一年生茎段作为外植体,考察不同的灭菌方法和激素配比对外植体灭菌、启动、增殖、生根的影响。结果表明:悬铃木茎段的最佳灭菌方法是75%乙醇灭菌30 s+0.1%HgCl2消毒17 min... 为了优化建立悬铃木组织培养再生体系,本研究以健壮、无病虫害的一年生茎段作为外植体,考察不同的灭菌方法和激素配比对外植体灭菌、启动、增殖、生根的影响。结果表明:悬铃木茎段的最佳灭菌方法是75%乙醇灭菌30 s+0.1%HgCl2消毒17 min,污染率可降至20%;最适宜的启动培养基为MS+ZT 4 mg/L+IAA 1.0 mg/L,启动率可达到65%;最适宜的增殖培养基为MS+ZT 3 mg/L+IAA 1.0 mg/L,增殖系数可达到1.95;最适宜的生根培养基为1/2 MS+IBA 0.1 mg/L,生根率可达到75%。 展开更多
关键词 悬铃木 组织培养 外植体 再生体系
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Optimal transcorneal electrical stimulation parameters for preserving photoreceptors in a mouse model of retinitis pigmentosa
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作者 Sam Enayati Karen Chang +10 位作者 Anton Lennikov Menglu Yang Cherin Lee Ajay Ashok Farris Elzaridi Christina Yen Kasim Gunes Jia Xie Kin-Sang Cho Tor Paaske Utheim Dong Feng Chen 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第11期2543-2552,共10页
Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on pho... Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on photoreceptor survival.This study aims to identify the most effective electrical stimulation parameters and functional advantages of transcorneal electrical stimulation(tcES)in mice affected by inherited retinal degeneration.Additionally,the study seeked to analyze the electric field that reaches the retina in both eyes in mice and post-mortem humans.In this study,we recorded waveforms and voltages directed to the retina during transcorneal electrical stimulation in C57BL/6J mice using an intraocular needle probe with rectangular,sine,and ramp waveforms.To investigate the functional effects of electrical stimulation on photoreceptors,we used human retinal explant cultures and rhodopsin knockout(Rho^(-/-))mice,demonstrating progressive photoreceptor degeneration with age.Human retinal explants isolated from the donors’eyes were then subjected to electrical stimulation and cultured for 48 hours to simulate the neurodegenerative environment in vitro.Photoreceptor density was evaluated by rhodopsin immunolabeling.In vivo Rho^(-/-)mice were subjected to two 5-day series of daily transcorneal electrical stimulation using rectangular and ramp waveforms.Retinal function and visual perception of mice were evaluated by electroretinography and optomotor response(OMR),respectively.Immunolabeling was used to assess the morphological and biochemical changes of the photoreceptor and bipolar cells in mouse retinas.Oscilloscope recordings indicated effective delivery of rectangular,sine,and ramp waveforms to the retina by transcorneal electrical stimulation,of which the ramp waveform required the lowest voltage.Evaluation of the total conductive resistance of the post-mortem human compared to the mouse eyes indicated higher cornea-to-retina resistance in human eyes.The temperature recordings during and after electrical stimulation indicated no significant temperature change in vivo and only a subtle temperature increase in vitro(~0.5-1.5°C).Electrical stimulation increased photoreceptor survival in human retinal explant cultures,particularly at the ramp waveform.Transcorneal electrical stimulation(rectangular+ramp)waveforms significantly improved the survival and function of S and M-cones and enhanced visual acuity based on the optomotor response results.Histology and immunolabeling demonstrated increased photoreceptor survival,improved outer nuclear layer thickness,and increased bipolar cell sprouting in Rho^(-/-)mice.These results indicate that transcorneal electrical stimulation effectively delivers the electrical field to the retina,improves photoreceptor survival in both human and mouse retinas,and increases visual function in Rho^(-/-)mice.Combined rectangular and ramp waveform stimulation can promote photoreceptor survival in a minimally invasive fashion. 展开更多
关键词 bipolar cells electrical stimulation NEUROPROTECTION photoreceptor degeneration RETINA retinal explants retinitis pigmentosa transcorneal electrical stimulation WAVEFORM
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Low-molecular-weight fucoidan inhibits the proliferation of melanoma via Bcl-2 phosphorylation and PTEN/AKT pathway
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作者 MINJI PARK CHULHWAN BANG +1 位作者 WON-SOO YUN YUN-MI JEONG 《Oncology Research》 SCIE 2024年第2期273-282,共10页
Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-He... Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-Helicobacter pylori properties.However,the effects of low-molecular-weight fucoidan(LMW-F)on melanoma cell lines and three dimensional(3D)cell culture models are not well understood.This study aimed to investigate the effects of LMW-F on A375 human melanoma cells and cryopreserved biospecimens derived from patients with advanced melanoma.Ultrasonic wave was used to fragment fucoidan derived from Fucus vesiculosus into smaller LMW-F.MTT and live/dead assays showed that LMW-F inhibited cell proliferation in both A375 cells and patientderived melanoma explants in a 3D-printed collagen scaffold.The PTEN/AKT pathway was found to be involved in the anti-melanoma effects of fucoidan.Western blot analysis revealed that LMW-F reduced the phosphorylation of Bcl-2 at Thr 56,which was associated with the prevention of anti-apoptotic activity of cancer cells.Our findings suggested that LMW-F could enhance anti-melanoma chemotherapy and improve the outcomes of patients with melanoma resistance. 展开更多
关键词 Low-molecular-weight fucoidan MELANOMA Patient-derived melanoma explants in a 3D-printed collagen scaffold Anti-melanoma effect PTEN-AKT-Bcl-2 network
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基于茎段外植体的茶树组培苗快繁技术研究
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作者 李家兴 朱俊彦 +6 位作者 张有泽 朱木兰 陈泓蓉 任露露 刘升锐 姚明哲 韦朝领 《茶叶学报》 2024年第1期28-36,共9页
【目的】目前,茶树组培苗繁育存在诱导出芽效率不高、繁殖系数较低等问题。因此,为提高茶树种质资源保存繁育效率,丰富保存手段与技术,建立一种高效、快速、实用的基于茎段外植体的茶树组织培养高效快繁技术体系非常必要。【方法】选用... 【目的】目前,茶树组培苗繁育存在诱导出芽效率不高、繁殖系数较低等问题。因此,为提高茶树种质资源保存繁育效率,丰富保存手段与技术,建立一种高效、快速、实用的基于茎段外植体的茶树组织培养高效快繁技术体系非常必要。【方法】选用来自安徽省六安市舒城县鲍家村群体种的一个国家级品种‘报春1号’作为研究试材,对该品种进行组织培养快繁技术的探索,包括消毒、定芽诱导、不定芽诱导(I期和II期)、芽伸长、以及生根等阶段。【结果】离体快繁:最佳定芽诱导培养基为MS+3 mg·L^(-1)6-苄基腺嘌呤(6-BA),诱导率达82.2%;不定芽增殖I期最佳培养基为MS基础培养基(MS)+2 mg·L^(-1)6-BA+0.2 mg·L^(-1)萘乙酸(NAA),诱导率为77.78%,平均芽数为5.4个;不定芽增殖II期最佳培养基为MS+1 mg·L^(-1)6-BA+0.3 mg·L^(-1)NAA+2 mg·L^(-1)噻苯隆(TDZ),诱导芽数最多(平均值为10.43个),芽苗呈绿色,叶片舒展;不定芽伸长最佳培养基为MS+0.8 mg·L^(-1)6-BA+0.08 mg·L^(-1)NAA,伸长率为63.33%;不定根诱导培养基以1/2MS培养基+3 mg·L^(-1)IBA效果较佳。【结论】本研究结果建立了一种基于茎段外植体的茶树组织培养高效快繁技术,将对优良茶树品种的保存繁育及其高效利用提供理论基础。 展开更多
关键词 茶树组培苗 组织培养 茎段外植体 高效快繁
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金柑组织培养与快速繁殖体系的建立
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作者 高照祖 黄红梅 《现代农业科技》 2024年第1期112-117,共6页
为了建立金柑组织培养与快速繁殖体系,以金柑胚根、子叶、茎段、叶片等不同外植体为材料,采用器官发生型以及短枝扦插型为主要培养途径,进行金柑组织培养,研究不同浓度植物生长调节剂对金柑快速繁殖体系建立的影响以及愈伤组织诱导和分... 为了建立金柑组织培养与快速繁殖体系,以金柑胚根、子叶、茎段、叶片等不同外植体为材料,采用器官发生型以及短枝扦插型为主要培养途径,进行金柑组织培养,研究不同浓度植物生长调节剂对金柑快速繁殖体系建立的影响以及愈伤组织诱导和分化的最适条件,筛选出诱导愈伤组织形成不定芽的最佳外植体与最适培养基。结果表明,金柑诱导愈伤组织形成不定芽的最佳外植体为幼嫩的茎段,诱导金柑子叶形成愈伤组织的最佳培养基为MS+6-BA 1.5 mg/L+NAA 1.0 mg/L+3%蔗糖+0.7%琼脂,诱导子叶愈伤组织分化不定芽的最佳培养基为MS+6-BA 1.0 mg/L+IBA 0.1 mg/L+3%蔗糖+0.7%琼脂。 展开更多
关键词 金柑 组织培养 快速繁殖 外植体
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兔膝骨关节炎成纤维样滑膜细胞分离培养方法的研究
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作者 陈芳 朱以良 +4 位作者 张传成 刘琴 喻晶 杨丽君 张宜 《生物骨科材料与临床研究》 CAS 2024年第1期14-18,25,共6页
目的在传统细胞组织块培养方法的基础上进行改进,建立一种简单、可重复性操作的兔膝骨关节炎成纤维样滑膜细胞分离培养方法。方法通过手术诱导建立兔膝骨关节炎模型,在无菌条件下,快速取出兔膝骨关节炎的滑膜组织,经过冲洗处理后剪成一... 目的在传统细胞组织块培养方法的基础上进行改进,建立一种简单、可重复性操作的兔膝骨关节炎成纤维样滑膜细胞分离培养方法。方法通过手术诱导建立兔膝骨关节炎模型,在无菌条件下,快速取出兔膝骨关节炎的滑膜组织,经过冲洗处理后剪成一定大小碎片,用无菌滤纸吸净组织上附着的水分,接种在培养皿后,进行观察、换液、传代、拍照。将原代培养的细胞传至第三代细胞接种于96孔板中,采用噻唑蓝(MTT)法分别在不同时间梯度检测各孔对应吸光度,制作生长曲线;采用免疫荧光染色法观察细胞中波形蛋白的表达情况。结果培养的细胞为典型的梭形,符合成纤维样滑膜细胞的生长形态特征,免疫荧光染色显示波形蛋白强表达。结论改进后的组织块培养方法简单、操作可重复性强,可以获得大量兔膝骨关节炎成纤维样滑膜细胞,为后续膝骨关节炎的机理研究提供了大量细胞支持。 展开更多
关键词 膝骨关节炎 成纤维样滑膜细胞 改良组织块法
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Investigation on the Morphological Characteristics of Dendrobium officinale Plantlets Propagated from Different Explants 被引量:5
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作者 Jiaxian LIANG Junjie MO +1 位作者 Hancai CAO Qianting TAN 《Agricultural Biotechnology》 CAS 2014年第5期11-14,20,共5页
[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets pro... [ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets propagated via different regeneration pathways were selected for morphological investigation and classification. [ Result] D. officinale plantlets propagated from stem segment explants exhibited highly consistent morphological characteristics, while those propagated from seed explants exhibited a variety of morphological characteristics. [ Conclusion] Therefore, using seed explants for regeneration can effectively broaden the germplasms resources of D. officinale. 展开更多
关键词 Dendrobium officinale Stem segment explants Seed explants Germplasms resources
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Establishment of high frequency shoot regeneration system in Himalayan poplar(Populus ciliata Wall. ex Royle) from petiole explants using Thidiazuron cytokinin as plant growth regulator 被引量:4
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作者 G. Aggarwal A. Gaur D. K. Srivastava 《Journal of Forestry Research》 SCIE CAS CSCD 2015年第3期651-656,共6页
Populus species are important resources for industry and in scientific study on biological and agricul- tural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar (Populus cil... Populus species are important resources for industry and in scientific study on biological and agricul- tural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar (Populus ciliata wall. ex Royle). The effect of TDZ alone and in combi- nation with adenine and NAA was studied on the regen- eration potential of petiole explants. The explants were excised from Himalayan poplar plants grown in glass- houses. After surface sterilization the explants were cul- tured on shoot induction medium. High percentage shoot regeneration (86 %) was recorded on MS medium sup- plemented with 0.004 mg L-1 TDZ and 79.7 mg L-1 adenine. The regenerated shoots for elongation and multi- plication were transferred to MS + 0.5 mg L-1 BAP + 0.2 mg L-1 IAA + 0.3 mg L-1 GA3. Root re- generation from shoots developed in vitro was observed on MS medium supplemented with 0.10 mg L-1 IBA. Hi- malayan poplar plantlets could be produced within 2 months after acclimatization in a sterile mixture of sand and soil. We developed a high efficiency plant regeneration protocol from petiole explants of P. ciliata. 展开更多
关键词 In vitro regeneration Petiole explants Growth regulator THIDIAZURON Populus ciliata
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Differentiation potential of human adipose tissue derived stem cells into photoreceptors through explants culture and enzyme methods 被引量:3
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作者 Wei-Wei Xu Li Huang +5 位作者 Kelvin K.L.Chong Doreen S.Y.Leung Benjamin EL.Li Zheng-Qin Yin Yi-Fei Huang Chi Pui Pang 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2017年第1期23-29,共7页
AIM: To investigate the retinal photoreceptor differentiation potential of human orbital adipose tissue-derived stem cells (ADSCs) generated by enzyme (EN) and explant (EX) culture methods.METHODS: We investig... AIM: To investigate the retinal photoreceptor differentiation potential of human orbital adipose tissue-derived stem cells (ADSCs) generated by enzyme (EN) and explant (EX) culture methods.METHODS: We investigated potentials of human orbital ADSCs to differentiate into photoreceptors through EN and EX culture methods. EN and EX orbital ADSCs were obtained from the same donor during rehabilitative orbital decompression, and then were subject to a 3-step induction using Noggin, DKK-1, IGF-1 and b-FGF at different time points for 38d. Stem cell, eye-field and photoreceptor-related gene and protein markers were measured by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescent (IMF) staining.RESULTS: Both EX and EN orbital ADSCs expressed CD133, a marker of cell differentiation. Moreover, PAX6 and rhodopsin, markers of the retinal progenitor cells, were detected from EX and EN orbital ADSCs. In EX orbital ADSCs, PAX6 mRNA was detected on the 17th day and then the rhodopsin mRNA was detected on the 24th day. In contrast, the EN orbital ADSCs expressed PAX6 and rhodopsin mRNA on the 31st day. EX orbital ADSCs expressed rhodopsin protein on the 24th day, while EN orbital ADSCs expressed rhodopsin protein on the 31st day. CONCLUSION: Orbital ADSCs isolated by direct explants culture show earlier and stronger expressions of markers towards eye field and retinal photoreceptor differentiation than those generated by conventional EN method. 展开更多
关键词 photoreceptor cells cell differentiation adultstem cells tissue engineering explants culture enzymaticdigestion
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An Improved System for Shoot Regeneration from Stem Explants of Lombardy Poplar (Populus nigra L. var. italica Koehne) 被引量:4
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作者 Kamal Kanti Biswas Takeshi Mohri +2 位作者 Satoshi Kogawara Yoshihiro Hase Yutaka Oono 《American Journal of Plant Sciences》 2012年第9期1181-1186,共6页
We developed a system for the regeneration of Lombardy poplar (Populus nigra L. var. italica) shoots from internodal stem explants. Using this system, shoots regenerated from 87% of the stem explants placed on Murashi... We developed a system for the regeneration of Lombardy poplar (Populus nigra L. var. italica) shoots from internodal stem explants. Using this system, shoots regenerated from 87% of the stem explants placed on Murashige and Skoog (MS) medium supplemented with 0.1 mg/L indole-3-acetic acid and 0.5 mg/L benzylaminopurine without undergoing callus formation. About 80% of the in vitro regenerated shoots developed roots on MS medium supplemented with 0.5 mg/L indole-3-butyric acid and 0.02 mg/L 1-naphthylacetic acid. Well-rooted seven-to eight-week-old regenerated plants could be transferred to soil for further growth and the survival rate of such plants after three weeks was 88%. The protocol presented here is simple and economical because it does not rely on pre-incubation in callus induction medium or repeated subculture in shoot induction medium containing trans-zeatin, an expensive substance. The in vitro regeneration system presented here could be used for evaluation of radiation sensitivity for Lombardy poplar tissues. 展开更多
关键词 Lombardy POPLAR SHOOT Regeneration STEM explantS AUXIN and Benzylaminopurine Radiation Sensitivity
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The Influence of Endogenous Hormones on Culture Capability of Different Explants in Rice 被引量:2
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作者 WANGXiu-hong SHIXiang-yuan +2 位作者 WUXian-jun WANGXu-dong ZHOUKai-da 《Agricultural Sciences in China》 CAS CSCD 2005年第5期343-347,共5页
The endogenous hormones (EHs) content of different explants (anther, young panicle, young embryo and mature embryo) and calli with different culture capability were analyzed by means of high performance liquid chromat... The endogenous hormones (EHs) content of different explants (anther, young panicle, young embryo and mature embryo) and calli with different culture capability were analyzed by means of high performance liquid chromatography (HPLC). The results showed that the contents and ratio of endogenous hormones were one of the key factors affecting callus induction frequencies (CIF) and green plantlet differentiation frequencies (GPDF). The influence of endogenous hormones of different explants on CIF represented as: Zoatin ribosile (ZR) showed negative effect, indole-3-acetic acid (IAA) did positive effect, and gibberellic acid (GA) did negative effect except for mature embryos. The influence of endogenous hormones on green plantlet differentiation frequency (GPDF) showed: IAA and GA were negative effect; abscisic acid (ABA) and zeatin+ zeatin riboside (Z+ZR) were positive effect. The mixture ratio of endogenous hormones played a role on CIF and GPDF. IAA/Z+ZR had a positive effect on CIF, and there was a notable positive correlation between Z+ZR/ IAA and GPDF, so was between ABA/IAA and GPDF. 展开更多
关键词 RICE Endogenous hormones (EHs) explant Callus induction frequency (CIF) Green plantlet differentiation frequency (GPDF)
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A pilot study of intraocular lens explantation in 69 eyes in Chinese patients 被引量:1
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作者 Fang Chai Bo Ma +2 位作者 Xin-Guang Yang Juan Li Mei-Fang Chu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2017年第4期579-585,共7页
AIM:To study the effects of intraocular lens(IOL) explantation and demographic characteristics.METHODS:Retrospective non-comparative case series.Clinical data recorded from patient charts included the following:d... AIM:To study the effects of intraocular lens(IOL) explantation and demographic characteristics.METHODS:Retrospective non-comparative case series.Clinical data recorded from patient charts included the following:demographic,preoperative and postoperative characteristics;complications;surgical methods,and changes in visual acuity.RESULTS:A total of 69 eyes in 67 Chinese patients who received IOL explants were studied.The patients' mean age at the time of explantation was 46.1 years old [SD 22.5(6-85)],and 37 patients were female(55.2%).Regarding employment,47.8% were farmers,23.9% were retired,16.4% were students,4.5% were unemployed,3% were workers,and 4.5% were other(including staff members,teachers and officers).The main reasons for explantation were dislocation/decentration in 41 cases(59.4%) and retinal detachment in 10 cases(14.5%).The third most prevalent cause was incorrect lens power in 7 eyes(10.1%).The remaining reasons were endophthalmitis in 6 cases(8.7%),posterior capsular opacity in 3 eyes(4.3%),and impacting retinal surgery operation in 2 cases(2.9%).The main comorbidities were high myopia in 18 eyes(26.1%),trauma in 8 eyes(11.6%),retinal detachment in 6 eyes(8.7%),congenital cataracts in 8 eyes(11.6%),and Marfan's syndrome in 2 eyes(2.9%).The mean time from implantation to explantation was 4.0y [SD 4.2(0.005-15)].Treatment after explantation included posterior chamber IOL implantation in 44 eyes(63.8%) and aphakia in 25 eyes(36.2%).After surgery,the best corrected visual ability(BCVA) was improved in 50 cases(72.5%),including 28 patients(40.6%) in whom visual ability was improved by more than two lines.CONCLUSION:Dislocation/decentration is the main cause for explantation,and high myopia is a main risk factor.Posterior chamber IOL implantation remains the most elected treatment after explantation. 展开更多
关键词 explantATION intraocular lens DISLOCATION
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Tissue Culture Responses from Different Explants of Rice 被引量:1
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作者 WANG Xiu-hong SHI Xiang-yuan Wu Xian-jun 《Rice science》 SCIE 2005年第3期229-232,共4页
关键词 RICE explant callus induction frequency green plantlet differentiation frequency CORRELATION
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