三七组织培养的建立

三七的根、块根、根茎、茎、叶柄、叶和花蕾等各部位诱导出愈伤组织。当这些愈伤组织无性系继代培养到第5代后进行了药用成分皂苷薄层层析的初步鉴定。结果表明三七愈伤组织中含有皂苷的主要成分Rg₁ Rb₁和Rh₁。粗总皂昔含量比原植物高,为干重的5．37％(原植物为干重的4．25％),粗皂苷元含量也高于原植物。愈伤组织的生长速率是54．0mg干重／L／day。各部位愈伤组织中以茎愈伤组织较好,无论干重增加、生长速率、粗总皂苷含量和产率、粗皂苷元含量还是皂甙组成上均比其他部位愈伤组织优越。对于茎愈伤组织的生长,基本培养基以Ms培养基较好,最适pH值为5．8,最适温度为26℃。光照对生长稍有促进但不显著。     

外源刺激物对长春花冠瘿细胞生长和吲哚生物碱含量的影响

所谓外源刺激物（ｅｌｉｃｉｔｏｒｓ）是指一些生物（多为真菌提取物）的或非生物的分子,它们能够通过信号传导途径,刺激植物发生防御反应,诱导特定的次级代谢产物的形成和积累^[1,5]。目前,有关elicitors对植物细胞次级代谢的影响和机制的研究引起了人们的普遍关注。我们曾经报道了以土壤农杆菌感染,Ti质粒转化得到的长春花冠瘿细胞,无论在生长还是在具有药用价值的次级代谢产物吲哚生物碱含量方面,都优于一般细胞培养常用的愈细胞^[2].本文研究了深红酵母匀浆物和果胶降解物作为外源刺激物,对长春花冠瘿细胞生长和吲哚生物碱含量的影响。     

不同理化因子对黄连愈伤组织生长和生物碱合成的影响

古蔺野连（又称串珠连Ｃｏｐｔｉｓｇｕｌｉｎｅｎｓｉｓ）为黄连属植物中的一个野生珍稀品种,只生长在我国西南少数山区,其根茎可作中药黄连,含有与商品黄连相同的小檗碱等化学成分^[1].作者在对黄连细胞培养研究中,通过目视法和TLC比较,从古蔺野连的愈伤组织中初步筛选到了生物量增长较快、生物碱合成能力较为稳定的无性系H292,进行了其生物量及生物碱含量的动态测定。本文报道植物激素、温度、光照等对黄连愈伤组织生长和生物碱合成能力的影响。     

裸燕麦胚性愈伤组织培养及悬浮系的建立

裸燕麦成熟胚在IN_６培养基上诱导培养,初始愈伤组织为白色、瘤状、外软内硬、易分化植株的非松脆类型。当在IM_１～IM₄培养基上进行循环式调控培养,经7～8个月,初始愈伤组织转变为浅黄色、小颗粒状、生活力强的松脆型胚性愈伤组织。将胚性愈伤组织置于液体培养基中悬浮培养,得到分散性好、生长快的悬浮细胞系。悬浮系经分化培养获得了再生植株,移栽成活率达95%以上。     

杜仲叶片愈伤组织诱导的激素优化研究

以杜仲优树L33的幼叶为材料,在B5培养基上添加不同浓度的生长素与细胞分裂素进行愈伤组织诱导研究,结果表明：无激素的培养基上不能诱导出愈伤组织,单独加入2,4-D、NAA和IBA均可诱导出愈伤组织,以0.5 mg·L^-1 2,4-D、0.5～1.0 mg·L^-1 NAA、1.0 mg·L^-1 IBA出愈率最高,达100%,且愈伤组织生长较好;在适宜浓度的生长素的培养基上加入细胞分裂素时,KT的加入对愈伤组织的诱导及生长起抑制作用;1.0 mg·L^-1 NAA+0.3～0.5 mg·L^-1 BA与1.0 mg·L^-1 IBA+0.3～0.5 mg·L^-1 BA 的组合可明显促进愈伤组织的诱导和生长,适合进一步继代培养。     

三裂叶野葛毛状根的诱导及其固体培养和液体培养

发根农杆菌（Agrobacterium rhizogenes）ATCC15834感染三裂叶野葛(Pueraria phaseoloides)叶片外植体20 d后产生毛状根,毛状根可直接从叶片外植体叶脉处或从叶脉处产生的愈伤组织上产生。感染35d后,约85%的叶片外植体产生毛状根。毛状根能在无外源生长调节剂的 MS固体和液体培养基上自主生长。PCR扩增结果表明,发根农杆菌Ri质粒的rolB和rolC基因已在三裂叶野葛毛状根基因组中整合并得到表达。与固体培养的毛状根相比,在液体培养基中培养的毛状根不仅生长迅速,也不会形成愈伤组织。在无外源生长调节剂的液体MS培养基中培养15d的三裂叶野葛毛状根的鲜重、干重、可溶性总糖含量及细胞内活性氧(ROS)含量分别为固体培养毛状根的1.59倍、1.18倍、5.25倍和1.16倍。     

灰绿黄堇愈伤组织培养及生物总碱含量的初步研究

将灰绿黄堇(Corydalis adunca Maxim．)无菌试管苗的茎段接种于附加不同BA和NAA浓度组合的MS培养基上,诱导出愈伤组织。实验表明较高浓度的BA和较低浓度的NAA组合有利于灰绿黄堇愈伤组织的诱导和生长以及愈伤组织中生物碱的合成。在附加BA2．00mg／L NAA0．2mg／L的MS培养基上,灰绿黄堇愈伤组织生长快,诱导率较高,合成的生物总碱含量较高,可达0．312％。     

人三叶因子3在毕赤酵母中表达条件的研究

为提高人三叶因子 3 (HumanTrefoilfactor 3 ,hTFF3 )在毕赤酵母中的表达量 ,研究了转化子生长的培养条件 ,包括不同碳源对转化子生长的影响和接种量、甲醇浓度、pH值、摇瓶转速及不同诱导时间对人三叶因子 3表达的影响。结果表明转化子在生长阶段加入葡萄糖生长旺盛 ,培养 14h后OD₆₀₀ 就可达到 50。在 100mL生长培养基上的菌液以 1∶1接入诱导培养基时蛋白表达量最高 ;转化子在 1%的甲醇、pH60、摇瓶转速240r/min的条件下诱导4 8h ,菌体密度OD₆₀₀为 15 ,目的蛋白表达量达到 20mg L。用 5L发酵罐进行了高密度发酵 ,经2%甲醇32h诱导 ,最终菌体密度OD₆₀₀ 达到 120 ,每升发酵液中含目的蛋白100mg。     

药用植物组织培养的研究 Ⅰ.三分三(Scopolia acutangula)愈伤组织的培养及莨菪碱、东莨菪碱的产生

本文研究了从三分三茎诱导愈伤组织的适宜培养基和该组织培养的适宜温度、pH 值和培养基、光、各种生长调节物质对其生长及莨菪碱、东莨菪碱含量的影响。应用纸层析、薄层层析和紫外光谱鉴定证明所获得的愈伤组织具有合成这两种生物碱的能力,并发现其在生物碱的组成和含量上与原茎不同,愈伤组织比原茎多3种化合物及一种显荧光的化合物。文中讨论了组织培养法用于工业生产的有关问题。     

云南红豆杉培养细胞系的建立

紫杉醇(Taxol)最初是从红豆杉属植物短叶红豆杉(Taxus brevifolia)树皮中分离出的一种二萜类化合物^[1]．对卵巢癌,转移性乳腺癌和恶性黑色寮瘤等患者疗效显著^[2],全世界红豆杉属植物有近11种,都含紫杉醇成分．但含量很低,加之现存数量很少,生长极为缓慢．造成了紫杉醇原料供应的危机^[3]。紫杉醇化学合成已经成功^[4-6],但繁杂的反应过程及前体化合物来源的限制使得它们无法实现商业化生产。最近从短叶红豆杉中分离出一种生产紫杉醇的内寄生真菌Tgromyer andreanae^[7]．由于紫杉醇含量仅为24～50ng/L．没有实用价值。植物细胞和组织培养可能是解决天然抗肿瘤药物长期供应的有效方法之一^[8]。自1991年Christen等人申请利用红豆杉细胞培养物生产紫杉醇专利以来^[9]．有关红豆杉细胞培养的研究已有不少报^[10-12]。但云南红豆杉(T.yunnanensis)仅见愈伤组织诱导的报道^[13]。本文报道云南红豆杉愈伤组织诱导和细胞培养的初步结果,并分析了细胞培养物中紫杉醇含量。     

颌下腺导管细胞与胶原海绵细胞相容性的实验研究

为探讨胶原海绵对颌下腺 (submandibulargland ,SMG)导管细胞的细胞相容性 ,采用HE染色光镜观察及免疫组化观察SMG导管细胞接种于胶原海绵后 ,细胞的生长情况。光镜下可见接种后第 1d细胞数量较少 ,分散于胶原海绵支架中间 ,第 7d细胞数量明显增加 ,免疫组织化学染色抗IV型胶原抗体染色呈阳性 ,说明细胞与支架材料之间已经有细胞外基质产生。胶原海绵具有良好的细胞相容性 ,是一种理想的支架材料。与胶原海绵复合培养 ,颌下腺导管细胞仍可保持良好的增殖能力。     

Determining Cell Number During Cell Culture using the Scepter Cell Counter

Counting cells is often a necessary but tedious step for in vitro cell culture. Consistent cell concentrations ensure experimental reproducibility and accuracy. Cell counts are important for monitoring cell health and proliferation rate, assessing immortalization or transformation, seeding cells for subsequent experiments, transfection or infection, and preparing for cell-based assays. It is important that cell counts be accurate, consistent, and fast, particularly for quantitative measurements of cellular responses.Despite this need for speed and accuracy in cell counting, 71% of 400 researchers surveyed¹ who count cells using a hemocytometer. While hemocytometry is inexpensive, it is laborious and subject to user bias and misuse, which results in inaccurate counts. Hemocytometers are made of special optical glass on which cell suspensions are loaded in specified volumes and counted under a microscope. Sources of errors in hemocytometry include: uneven cell distribution in the sample, too many or too few cells in the sample, subjective decisions as to whether a given cell falls within the defined counting area, contamination of the hemocytometer, user-to-user variation, and variation of hemocytometer filling rate².To alleviate the tedium associated with manual counting, 29% of researchers count cells using automated cell counting devices; these include vision-based counters, systems that detect cells using the Coulter principle, or flow cytometry¹. For most researchers, the main barrier to using an automated system is the price associated with these large benchtop instruments¹.The Scepter cell counter is an automated handheld device that offers the automation and accuracy of Coulter counting at a relatively low cost. The system employs the Coulter principle of impedance-based particle detection³ in a miniaturized format using a combination of analog and digital hardware for sensing, signal processing, data storage, and graphical display. The disposable tip is engineered with a microfabricated, cell- sensing zone that enables discrimination by cell size and cell volume at sub-micron and sub-picoliter resolution. Enhanced with precision liquid-handling channels and electronics, the Scepter cell counter reports cell population statistics graphically displayed as a histogram.     

细胞提取物重编程研究进展

体细胞重编程是在特定的条件下使已分化的细胞转变成为另一种细胞.体细胞重编程的方式主要有体细胞核移植技术、细胞融合技术、细胞提取物处理技术及特定转录因子转染技术.现有研究表明,细胞提取物重编程技术在体细胞重编程中发挥着一定的作用,为此,就该技术的最新研究进展和可能机制作一综述.     

Cell Physician: Reading Cell Motion

Cell motility is an essential phenomenon in almost all living organisms. It is natural to think that behavioral or shape changes of a cell bear information about the underlying mechanisms that generate these changes. Reading cell motion, namely, understanding the underlying biophysical and mechanochemical processes, is of paramount importance. The mathematical model developed in this paper determines some physical features and material properties of the cells locally through analysis of live cell image sequences and uses this information to make further inferences about the molecular structures, dynamics, and processes within the cells, such as the actin network, microdomains, chemotaxis, adhesion, and retrograde flow. The generality of the principals used in formation of the model ensures its wide applicability to different phenomena at various levels. Based on the model outcomes, we hypothesize a novel biological model for collective biomechanical and molecular mechanism of cell motion.     

Tuning Cell Motility via Cell Tension with a Mechanochemical Cell Migration Model

Cell migration is orchestrated by a complicated mechanochemical system. However, few cell migration models take into account the coupling between the biochemical network and mechanical factors. Here, we construct a mechanochemical cell migration model to study the cell tension effect on cell migration. Our model incorporates the interactions between Rac-GTP, Rac-GDP, F-actin, myosin, and cell tension, and it is very convenient in capturing the change of cell shape by taking the phase field approach. This model captures the characteristic features of cell polarization, cell shape change, and cell migration modes. It shows that cell tension inhibits migration ability monotonically when cells are applied with persistent external stimuli. On the other hand, if random internal noise is significant, the regulation of cell tension exerts a nonmonotonic effect on cell migration. Because the increase of cell tension hinders the formation of multiple protrusions, migration ability could be maximized at intermediate cell tension under random internal noise. These model predictions are consistent with our single-cell experiments and other experimental results.