首页 > 最新文献

In Vitro最新文献

英文 中文
Lymphatic endothelial and smooth-muscle cells in tissue culture. 组织培养中的淋巴内皮细胞和平滑肌细胞。
Pub Date : 1984-07-01 DOI: 10.1007/BF02639772
M G Johnston, M A Walker

Endothelial and smooth-muscle cells from bovine mesenteric lymphatic vessels have been collected and cultured in vitro. The endothelial cells grew as a monolayer exhibiting a "cobblestone" appearance with individual cells tending to be more flattened at confluence than their blood vascular counterparts. Approximately 30% of these cells expressed Factor VIII antigen compared with bovine mesenteric artery or human umbilical-vein endothelium in which the majority of cells were positive. The lymphatic smooth-muscle cells exhibited focal areas of multilayering and were Factor VIII negative. The availability of lymphatic endothelial and smooth-muscle cells in culture will provide a new tool for the investigation of the biological properties of the lymphatic vessels and their role in homeostasis.

从牛肠系膜淋巴管中收集并体外培养内皮细胞和平滑肌细胞。内皮细胞呈单层生长,呈“鹅卵石”状,单个细胞在汇合处比血管细胞更扁平。这些细胞中约30%表达因子VIII抗原,而牛肠系膜动脉或人脐静脉内皮细胞中大部分细胞呈阳性。淋巴平滑肌细胞呈多层灶状,因子VIII阴性。淋巴内皮细胞和平滑肌细胞的培养将为研究淋巴管的生物学特性及其在体内平衡中的作用提供新的工具。
{"title":"Lymphatic endothelial and smooth-muscle cells in tissue culture.","authors":"M G Johnston,&nbsp;M A Walker","doi":"10.1007/BF02639772","DOIUrl":"https://doi.org/10.1007/BF02639772","url":null,"abstract":"<p><p>Endothelial and smooth-muscle cells from bovine mesenteric lymphatic vessels have been collected and cultured in vitro. The endothelial cells grew as a monolayer exhibiting a \"cobblestone\" appearance with individual cells tending to be more flattened at confluence than their blood vascular counterparts. Approximately 30% of these cells expressed Factor VIII antigen compared with bovine mesenteric artery or human umbilical-vein endothelium in which the majority of cells were positive. The lymphatic smooth-muscle cells exhibited focal areas of multilayering and were Factor VIII negative. The availability of lymphatic endothelial and smooth-muscle cells in culture will provide a new tool for the investigation of the biological properties of the lymphatic vessels and their role in homeostasis.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"566-72"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639772","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17491097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 24
Substratum attachment of embryonic mesoderm cells in culture. 胚胎中胚层细胞的基质附着。
Pub Date : 1984-07-01 DOI: 10.1007/BF02639767
E J Sanders

The cell-substratum adhesive characteristics of cultured chick embryo primary mesoderm cells have been examined by interference reflection microscopy and transmission electron microscopy under various conditions. Correlations were drawn between the type of adhesion and the degree of motility shown by the cells. During the rapid spreading and motility of cells cultured on fibronectin-containing substrata, focal contacts (10 to 15-nm gap) were rare and close contacts (about 30-nm gap) were predominant. By contrast, when the cells were immobile, after 5 d in culture, extensive focal contacts were present, together with stress fibers. The results indicate that tight cell-substratum contact is incompatible with rapid cell motility and that fibronectin acts by inducing the formation of close contacts rather than focal contacts.

采用干涉反射显微镜和透射电镜观察了不同条件下培养鸡胚初代中胚层细胞的细胞-基质黏附特性。绘制了粘附类型与细胞运动程度之间的相关性。在含纤维连接蛋白基质上培养的细胞快速扩散和运动过程中,局灶性接触(10 ~ 15nm间隙)很少,密切接触(约30nm间隙)居多。相比之下,当细胞不移动时,培养5 d后,存在广泛的局灶接触,以及应力纤维。结果表明,紧密的细胞-基质接触与快速的细胞运动不相容,纤维连接蛋白通过诱导密切接触而不是局灶接触而起作用。
{"title":"Substratum attachment of embryonic mesoderm cells in culture.","authors":"E J Sanders","doi":"10.1007/BF02639767","DOIUrl":"https://doi.org/10.1007/BF02639767","url":null,"abstract":"<p><p>The cell-substratum adhesive characteristics of cultured chick embryo primary mesoderm cells have been examined by interference reflection microscopy and transmission electron microscopy under various conditions. Correlations were drawn between the type of adhesion and the degree of motility shown by the cells. During the rapid spreading and motility of cells cultured on fibronectin-containing substrata, focal contacts (10 to 15-nm gap) were rare and close contacts (about 30-nm gap) were predominant. By contrast, when the cells were immobile, after 5 d in culture, extensive focal contacts were present, together with stress fibers. The results indicate that tight cell-substratum contact is incompatible with rapid cell motility and that fibronectin acts by inducing the formation of close contacts rather than focal contacts.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"521-7"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639767","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17526763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Properties of peroxisomes and their induction by clofibrate in normal adult rat hepatocytes in primary culture. 原代培养正常成年大鼠肝细胞中过氧化物酶体的特性及氯贝特对其的诱导作用。
Pub Date : 1984-07-01 DOI: 10.1007/BF02639773
K Furukawa, Y Mochizuki, N Sawada

Alterations in peroxisomes and catalase activity and their responsiveness to clofibrate in adult rat hepatocytes in primary culture were investigated. The numbers of peroxisomes with and without crystalloid nucleotids per unit cytoplasmic area were preserved in cultured hepatocytes for 2 d after seeding at a level comparable to that of freshly isolated hepatocytes. At Day 3 in culture, the number of anucleoid peroxisomes was reduced in untreated hepatocytes, accompanied by more significant reduction in the number of nucleoid-containing peroxisomes, which decreased until Day 5. Peroxisome diameters were reduced in untreated hepatocytes at Day 2 and this decrease in the diameter was continued until Day 7. Catalase activity in untreated hepatocytes decreased markedly with culture age. The number of anucleoid peroxisomes was significantly greater in hepatocytes treated with 2 mM clofibrate in culture than in freshly isolated hepatocytes for 2 d or in untreated hepatocytes of the same culture age through 7 d. The number of nucleoid-containing peroxisomes in the treated cells began to decrease in 3 d, but was greater than that of untreated cells at Days 3 and 5. Peroxisomes with well-developed nucleoids were observed frequently in the treated cells even at Day 7. Peroxisome diameters were greater in the treated cells than in untreated cells at Days 3, 5, and 7. Catalase activity was always higher in the treated cells than in untreated cells. These results suggest that clofibrate is effective in inducing peroxisome proliferation as well as in maintaining the organelles in cultured hepatocytes.

研究了原代培养的成年大鼠肝细胞中过氧化物酶体和过氧化氢酶活性的变化及其对氯贝特的反应。在培养的肝细胞中,每单位细胞质面积含有和不含晶体核苷的过氧化物酶体数量在播种后保存2天,其水平与新鲜分离的肝细胞相当。在培养第3天,未经处理的肝细胞中无核样过氧化物酶体的数量减少,同时含核过氧化物酶体的数量减少更为显著,直到第5天减少。未经处理的肝细胞在第2天过氧化物酶体直径减小,这种减小持续到第7天。未经处理的肝细胞过氧化氢酶活性随培养年龄明显降低。与新鲜分离的肝细胞相比,经2 mM克罗贝特处理2天的肝细胞中无核样过氧化物酶体的数量显著增加,或在相同培养年龄的肝细胞中未经处理7天。处理细胞中含核过氧化物酶体的数量在第3天开始减少,但在第3天和第5天高于未经处理的细胞。即使在第7天,在处理的细胞中也经常观察到具有发育良好的类核的过氧化物酶体。在第3、5和7天,处理过的细胞的过氧化物酶体直径大于未处理的细胞。过氧化氢酶活性在处理过的细胞中始终高于未处理的细胞。这些结果表明,氯贝酸钠在诱导过氧化物酶体增殖和维持培养肝细胞细胞器方面是有效的。
{"title":"Properties of peroxisomes and their induction by clofibrate in normal adult rat hepatocytes in primary culture.","authors":"K Furukawa,&nbsp;Y Mochizuki,&nbsp;N Sawada","doi":"10.1007/BF02639773","DOIUrl":"https://doi.org/10.1007/BF02639773","url":null,"abstract":"<p><p>Alterations in peroxisomes and catalase activity and their responsiveness to clofibrate in adult rat hepatocytes in primary culture were investigated. The numbers of peroxisomes with and without crystalloid nucleotids per unit cytoplasmic area were preserved in cultured hepatocytes for 2 d after seeding at a level comparable to that of freshly isolated hepatocytes. At Day 3 in culture, the number of anucleoid peroxisomes was reduced in untreated hepatocytes, accompanied by more significant reduction in the number of nucleoid-containing peroxisomes, which decreased until Day 5. Peroxisome diameters were reduced in untreated hepatocytes at Day 2 and this decrease in the diameter was continued until Day 7. Catalase activity in untreated hepatocytes decreased markedly with culture age. The number of anucleoid peroxisomes was significantly greater in hepatocytes treated with 2 mM clofibrate in culture than in freshly isolated hepatocytes for 2 d or in untreated hepatocytes of the same culture age through 7 d. The number of nucleoid-containing peroxisomes in the treated cells began to decrease in 3 d, but was greater than that of untreated cells at Days 3 and 5. Peroxisomes with well-developed nucleoids were observed frequently in the treated cells even at Day 7. Peroxisome diameters were greater in the treated cells than in untreated cells at Days 3, 5, and 7. Catalase activity was always higher in the treated cells than in untreated cells. These results suggest that clofibrate is effective in inducing peroxisome proliferation as well as in maintaining the organelles in cultured hepatocytes.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"573-84"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639773","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17528258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 3
Solute concentration effects on the expression of cellular heterogeneity of anchorage-independent growth among spontaneously transformed BALB/c3T3 cells. 溶质浓度对自发转化BALB/c3T3细胞非锚定生长细胞异质性表达的影响
Pub Date : 1984-07-01 DOI: 10.1007/BF02639774
H Rubin, B M Chu

Clones were derived in culture from a tumor initiated by spontaneously transformed 3T3 cells and tested for their colony-forming efficiency in agar (CFEag). Incubation of petri dish cultures was done in subsaturation humidity to minimize mold contamination. There was great variation in CFEag between clones but also, under certain conditions, within clones. The most prominent condition that generated phenotypic diversity in CFEag was partial evaporation of the medium, which may occur during the protracted development of a mass population from a single cell. Evaporation was disproportionately great in 35-mm dishes and peripheral wells of multiwell plates. If the supraphysiological solute concentration resulting from evaporation was greater than 133% of normal, there was progressive suppression of cell growth in the succeeding transfer in agar or on plastic, even if isotonic medium was substituted 1 d before transfer. The effect of supraphysiological concentrations of all the solutes of the medium could be reproduced by simply increasing the NaCl concentration. Damaged cells were restored to their full growth potential after 3 d in isotonic medium. When nontransformed cells were chronically exposed to increased salt, irreversible increases in 2-deoxyglucose uptake were produced. With continued exposure of these cells to high salt, they became morphologically transformed, produced colonies in agar with high efficiency, and formed sarcomas when inoculated into nude mice.

从一个由自发转化的3T3细胞引发的肿瘤中获得克隆,并在琼脂(CFEag)中测试其集落形成效率。培养皿培养在亚饱和湿度下进行,以尽量减少霉菌污染。无性系之间的CFEag存在很大差异,但在某些条件下,无性系内部也存在很大差异。在CFEag中产生表型多样性的最突出条件是培养基的部分蒸发,这可能发生在单细胞大规模群体的长期发育过程中。在多孔板的35mm培养皿和周边孔中,蒸发不成比例地大。如果蒸发产生的超生理溶质浓度大于正常的133%,则在琼脂或塑料上的后续转移中,即使在转移前1天替换等渗培养基,细胞生长也会逐渐受到抑制。培养基中所有溶质浓度的超生理效应都可以通过增加NaCl浓度来重现。损伤细胞在等渗培养基中培养3 d后恢复其完全生长潜能。当非转化细胞长期暴露于增加的盐中时,2-脱氧葡萄糖摄取产生不可逆的增加。随着这些细胞持续暴露于高盐环境中,它们发生形态转化,在琼脂中高效率地产生菌落,并在接种到裸鼠体内形成肉瘤。
{"title":"Solute concentration effects on the expression of cellular heterogeneity of anchorage-independent growth among spontaneously transformed BALB/c3T3 cells.","authors":"H Rubin,&nbsp;B M Chu","doi":"10.1007/BF02639774","DOIUrl":"https://doi.org/10.1007/BF02639774","url":null,"abstract":"<p><p>Clones were derived in culture from a tumor initiated by spontaneously transformed 3T3 cells and tested for their colony-forming efficiency in agar (CFEag). Incubation of petri dish cultures was done in subsaturation humidity to minimize mold contamination. There was great variation in CFEag between clones but also, under certain conditions, within clones. The most prominent condition that generated phenotypic diversity in CFEag was partial evaporation of the medium, which may occur during the protracted development of a mass population from a single cell. Evaporation was disproportionately great in 35-mm dishes and peripheral wells of multiwell plates. If the supraphysiological solute concentration resulting from evaporation was greater than 133% of normal, there was progressive suppression of cell growth in the succeeding transfer in agar or on plastic, even if isotonic medium was substituted 1 d before transfer. The effect of supraphysiological concentrations of all the solutes of the medium could be reproduced by simply increasing the NaCl concentration. Damaged cells were restored to their full growth potential after 3 d in isotonic medium. When nontransformed cells were chronically exposed to increased salt, irreversible increases in 2-deoxyglucose uptake were produced. With continued exposure of these cells to high salt, they became morphologically transformed, produced colonies in agar with high efficiency, and formed sarcomas when inoculated into nude mice.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"585-96"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639774","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17528257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 5
Porcine aortic organ culture: a model to study the cellular response to vascular injury. 猪主动脉器官培养:研究细胞对血管损伤反应的模型。
Pub Date : 1984-07-01 DOI: 10.1007/BF02639769
A I Gotlieb, P Boden

Organ cultures of porcine thoracic aorta were studied to define the characteristics of this system as a model to study the reaction of endothelial cells (ECs) and the underlying smooth muscle cells (SMCs) to injury. Both nonwounded and wounded cultures, the latter having had part of the endothelial surface gently denuded with a scalpel blade, were studied over a 7 d period by scanning and transmission electron microscopy. The results showed that the nonwounded ECs underwent a shape change from elongated to polygonal within 24 h in culture. In both nonwounded and wounded explants there was cell proliferation beneath the nondenuded endothelium so that by 7 d several layers of cells were present showing features of the secretory type of SMCs. This proliferation, however, did not occur if the endothelium was totally removed from the aorta. There was also evidence of gaps between the surface ECs, and by 7 d lamellipodia of cells beneath the surface were present in these gaps. Occasionally, elongated cells were seen to be present on the surface of the endothelium. In the wounded organ culture, cell migration and proliferation occurred extending from the wound edge and producing a covering of cells on the denuded area. There were also multilayered cells beneath the surface similar to the nonwounded area. Occasional foam cells were seen in the depth of the multilayered proliferating cells. The results indicate that organ culture of porcine thoracic aorta is a good model to study the reaction of ECs and underlying SMCs to injury.

通过对猪胸主动脉器官培养的研究,确定了该系统的特征,并以此为模型研究了内皮细胞(ECs)和下层平滑肌细胞(SMCs)对损伤的反应。在7天的时间里,通过扫描和透射电子显微镜研究非损伤和损伤培养,后者的部分内皮表面被手术刀刀片轻轻剥去。结果表明,未损伤的内皮细胞在培养24 h内由细长形变为多角形。未损伤和损伤的外植体在未剥落的内皮下都有细胞增殖,到第7天,出现了几层细胞,表现出SMCs的分泌型特征。然而,如果内皮完全从主动脉移除,则不会发生这种增殖。表面内皮细胞之间也有间隙,到第7天,这些间隙中出现了表面下的细胞板足。偶尔在内皮表面可见细长的细胞。在损伤器官培养中,细胞从创面边缘开始迁移和增殖,在剥落区形成细胞覆盖。表面下也有类似于未受伤区域的多层细胞。多层增生细胞深部偶见泡沫细胞。结果表明,猪胸主动脉的器官培养是研究ECs和下层SMCs对损伤反应的良好模型。
{"title":"Porcine aortic organ culture: a model to study the cellular response to vascular injury.","authors":"A I Gotlieb,&nbsp;P Boden","doi":"10.1007/BF02639769","DOIUrl":"https://doi.org/10.1007/BF02639769","url":null,"abstract":"<p><p>Organ cultures of porcine thoracic aorta were studied to define the characteristics of this system as a model to study the reaction of endothelial cells (ECs) and the underlying smooth muscle cells (SMCs) to injury. Both nonwounded and wounded cultures, the latter having had part of the endothelial surface gently denuded with a scalpel blade, were studied over a 7 d period by scanning and transmission electron microscopy. The results showed that the nonwounded ECs underwent a shape change from elongated to polygonal within 24 h in culture. In both nonwounded and wounded explants there was cell proliferation beneath the nondenuded endothelium so that by 7 d several layers of cells were present showing features of the secretory type of SMCs. This proliferation, however, did not occur if the endothelium was totally removed from the aorta. There was also evidence of gaps between the surface ECs, and by 7 d lamellipodia of cells beneath the surface were present in these gaps. Occasionally, elongated cells were seen to be present on the surface of the endothelium. In the wounded organ culture, cell migration and proliferation occurred extending from the wound edge and producing a covering of cells on the denuded area. There were also multilayered cells beneath the surface similar to the nonwounded area. Occasional foam cells were seen in the depth of the multilayered proliferating cells. The results indicate that organ culture of porcine thoracic aorta is a good model to study the reaction of ECs and underlying SMCs to injury.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"535-42"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639769","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17528254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 13
Effects of glucocorticoids and antiglucocorticoid on angiotensinogen production by hepatoma cells in culture. 糖皮质激素和抗糖皮质激素对培养肝癌细胞血管紧张素原产生的影响。
Pub Date : 1984-07-01 DOI: 10.1007/BF02639768
E Coezy, J Bouhnik, E Clauser, F Pinet, M Philippe, J Menard, P Corvol

Angiotensinogen is synthesized in large amounts by Fao cells derived from the Reuber H35 rat hepatoma in a medium enriched with 5% fetal bovine serum (FBS). Treatment of FBS with dextran-coated charcoal removed endogenous steroids without modifying angiotensinogen production. This treatment allowed the study of the effects of steroids on angiotensinogen production. Hydrocortisone increased the angiotensinogen synthesis in a dose-dependent manner. The antiglucocorticoid RU 38486 did not change the basal rate of angiotensinogen production but inhibited the stimulation by hydrocortisone. Similar results were obtained with dexamethasone. Angiotensinogen biosynthesis seems to be regulated by two distinct mechanisms: (a) glucocorticoid independent, controlling the basal rate of angiotensinogen production and (b) glucocorticoid dependent, mediating the increased rate of angiotensinogen production upon glucocorticoid treatment.

血管紧张素原是由来源于Reuber H35大鼠肝癌的Fao细胞在富含5%胎牛血清(FBS)的培养基中大量合成的。用葡聚糖包被木炭治疗FBS去除内源性类固醇而不改变血管紧张素原的产生。这种治疗方法使研究类固醇对血管紧张素原产生的影响成为可能。氢化可的松以剂量依赖性方式增加血管紧张素原的合成。抗糖皮质激素ru38486不改变血管紧张素原产生的基础速率,但抑制氢化可的松的刺激。地塞米松也获得了类似的结果。血管紧张素原的生物合成似乎受到两种不同机制的调节:(a)不依赖糖皮质激素,控制血管紧张素原产生的基础速率;(b)依赖糖皮质激素,介导糖皮质激素治疗后血管紧张素原产生速率的增加。
{"title":"Effects of glucocorticoids and antiglucocorticoid on angiotensinogen production by hepatoma cells in culture.","authors":"E Coezy,&nbsp;J Bouhnik,&nbsp;E Clauser,&nbsp;F Pinet,&nbsp;M Philippe,&nbsp;J Menard,&nbsp;P Corvol","doi":"10.1007/BF02639768","DOIUrl":"https://doi.org/10.1007/BF02639768","url":null,"abstract":"<p><p>Angiotensinogen is synthesized in large amounts by Fao cells derived from the Reuber H35 rat hepatoma in a medium enriched with 5% fetal bovine serum (FBS). Treatment of FBS with dextran-coated charcoal removed endogenous steroids without modifying angiotensinogen production. This treatment allowed the study of the effects of steroids on angiotensinogen production. Hydrocortisone increased the angiotensinogen synthesis in a dose-dependent manner. The antiglucocorticoid RU 38486 did not change the basal rate of angiotensinogen production but inhibited the stimulation by hydrocortisone. Similar results were obtained with dexamethasone. Angiotensinogen biosynthesis seems to be regulated by two distinct mechanisms: (a) glucocorticoid independent, controlling the basal rate of angiotensinogen production and (b) glucocorticoid dependent, mediating the increased rate of angiotensinogen production upon glucocorticoid treatment.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"528-34"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639768","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17528253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Establishment and characterization of cell lines from the Walker carcinoma 256 able to grow in suspension culture and deficient in thymidine kinase. 胸苷激酶缺失、悬浮培养的Walker癌256细胞系的建立与鉴定。
Pub Date : 1984-07-01 DOI: 10.1007/BF02639771
F Arvelo, A Yabrudi, M E Delgado, N González-Cadavid

A cell line from the Walker carcinosarcoma 256 of the rat has been established in suspension culture in medium with 5% bovine calf serum for over 350 generations, with an average population doubling time of 17 h, a plating efficiency of 56%, a colony forming efficiency of 32%, and a good capacity to form colonies in soft agar. The cells are morphologically indistinguishable from those in the solid tumor and ascites as checked by transmission and scanning electron microscopy. The karyotype is characterized by a modal number of 65 chromosomes and by the presence of a marker metacentric chromosome. The cells express thymidine kinase, gamma-glutamyl transpeptidase, and alkaline phosphatase; are agglutinable by concanavalin A; and can be synchronized by the triple thymidine block. They induce primary tumors, both subcutaneously (solid) and intraperitoneally (ascitic), in the rat; are able to metastasize upon injection by the tail vein; and invade the chorioallantoic membrane of the chick embryo. Cells in suspension can be transferred to monolayers, considerably decreasing their tumorigenicity without affecting the other parameters studied, and can be switched back to suspension culture. DNA-mediated transfection showed that DNA from these cells can transform the NIH-3T3 line. Upon growth of the monolayers in a BrdUr-containing medium, a sub-line was established that was cloned into a thymidine kinase-deficient line unable to grow in HAT medium and with properties otherwise similar to those of the parental wild type cells.

在含5%牛血清的培养基中悬浮培养大鼠Walker癌肉瘤256细胞系,培养350代以上,平均群体翻倍时间为17 h,培养效率为56%,菌落形成效率为32%,在软琼脂中具有良好的菌落形成能力。通过透射电镜和扫描电镜检查,这些细胞在形态上与实体瘤和腹水中的细胞难以区分。核型的特征是65条染色体的模态数和一个标记的异心染色体的存在。细胞表达胸苷激酶、γ -谷氨酰转肽酶和碱性磷酸酶;被豆豆蛋白A粘合;并且可以被三重胸腺嘧啶阻滞同步。它们诱导大鼠皮下(实体)和腹腔内(腹水)的原发性肿瘤;经尾静脉注射后能够转移;侵入鸡胚的绒毛膜尿囊膜。悬浮中的细胞可以转移到单层中,在不影响其他研究参数的情况下大大降低其致瘤性,并且可以切换回悬浮培养。DNA介导转染表明,来自这些细胞的DNA可以转化NIH-3T3细胞系。单层细胞在含有brdurd的培养基中生长后,建立了一个亚系,该亚系克隆到一个胸腺嘧啶激酶缺陷系中,该系不能在HAT培养基中生长,其特性与亲本野生型细胞相似。
{"title":"Establishment and characterization of cell lines from the Walker carcinoma 256 able to grow in suspension culture and deficient in thymidine kinase.","authors":"F Arvelo,&nbsp;A Yabrudi,&nbsp;M E Delgado,&nbsp;N González-Cadavid","doi":"10.1007/BF02639771","DOIUrl":"https://doi.org/10.1007/BF02639771","url":null,"abstract":"<p><p>A cell line from the Walker carcinosarcoma 256 of the rat has been established in suspension culture in medium with 5% bovine calf serum for over 350 generations, with an average population doubling time of 17 h, a plating efficiency of 56%, a colony forming efficiency of 32%, and a good capacity to form colonies in soft agar. The cells are morphologically indistinguishable from those in the solid tumor and ascites as checked by transmission and scanning electron microscopy. The karyotype is characterized by a modal number of 65 chromosomes and by the presence of a marker metacentric chromosome. The cells express thymidine kinase, gamma-glutamyl transpeptidase, and alkaline phosphatase; are agglutinable by concanavalin A; and can be synchronized by the triple thymidine block. They induce primary tumors, both subcutaneously (solid) and intraperitoneally (ascitic), in the rat; are able to metastasize upon injection by the tail vein; and invade the chorioallantoic membrane of the chick embryo. Cells in suspension can be transferred to monolayers, considerably decreasing their tumorigenicity without affecting the other parameters studied, and can be switched back to suspension culture. DNA-mediated transfection showed that DNA from these cells can transform the NIH-3T3 line. Upon growth of the monolayers in a BrdUr-containing medium, a sub-line was established that was cloned into a thymidine kinase-deficient line unable to grow in HAT medium and with properties otherwise similar to those of the parental wild type cells.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 7","pages":"549-65"},"PeriodicalIF":0.0,"publicationDate":"1984-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02639771","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17528256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Isolation and characterization of a near-diploid differentiated cell line from a murine teratocarcinoma that differentiates into muscle. 从小鼠畸胎瘤分化成肌肉的近二倍体分化细胞系的分离和特性。
Pub Date : 1984-06-01 DOI: 10.1007/BF02619619
E E Moore

Cell lines corresponding to various cell lineages of the mouse embryo have been isolated from murine teratocarcinomas. Embryonal carcinoma cell lines are developmentally equivalent to the embryonic ectoderm or inner cell mass. Most of these cell lines have a modal chromosome number equal or close to 40, the normal mouse complement. However, cell lines corresponding to more advanced cell lineages (e.g., endoderm) are tetraploid or hypotetraploid and display multiple chromosomal rearrangements. This paper describes the isolation of a near-diploid differentiated cell line (LT-D) from an LT teratocarcinoma. The modal chromosome number of LT-D is 40, and this number is stable during at least 12 mo of continuous culture. LT-D cells are morphologically distinct from embryonal carcinoma cells and no longer express the SSEA-1 cell surface antigen or high alkaline phosphatase activity characteristic of embryonal carcinoma cells. LT-D cells can be induced to fuse into structures resembling myotubes. The formation of these structures is accompanied by expression of the muscle-specific isozyme of creatine phosphokinase and desmin, a muscle-specific component of intermediate filaments. Lastly, LT-D cells do not form tumors in syngenetic mice.

从小鼠畸胎癌中分离出了与小鼠胚胎不同细胞系相对应的细胞系。胚胎癌细胞系在发育上等同于胚胎外胚层或内细胞团。大多数细胞系的模态染色体数目等于或接近正常小鼠补体的40。然而,与更高级的细胞系(例如,内胚层)相对应的细胞系是四倍体或亚四倍体,并显示多重染色体重排。本文报道了从LT畸胎癌中分离出的近二倍体分化细胞系(LT- d)。LT-D的模态染色体数为40,在至少12个月的连续培养中,这个数是稳定的。LT-D细胞在形态上与胚胎癌细胞不同,不再表达胚胎癌细胞所特有的SSEA-1细胞表面抗原或高碱性磷酸酶活性。LT-D细胞可以被诱导融合成类似肌管的结构。这些结构的形成伴随着肌酸磷酸激酶和desmin(中间纤维的肌肉特异性成分)的肌肉特异性同工酶的表达。最后,LT-D细胞在同基因小鼠中不会形成肿瘤。
{"title":"Isolation and characterization of a near-diploid differentiated cell line from a murine teratocarcinoma that differentiates into muscle.","authors":"E E Moore","doi":"10.1007/BF02619619","DOIUrl":"https://doi.org/10.1007/BF02619619","url":null,"abstract":"<p><p>Cell lines corresponding to various cell lineages of the mouse embryo have been isolated from murine teratocarcinomas. Embryonal carcinoma cell lines are developmentally equivalent to the embryonic ectoderm or inner cell mass. Most of these cell lines have a modal chromosome number equal or close to 40, the normal mouse complement. However, cell lines corresponding to more advanced cell lineages (e.g., endoderm) are tetraploid or hypotetraploid and display multiple chromosomal rearrangements. This paper describes the isolation of a near-diploid differentiated cell line (LT-D) from an LT teratocarcinoma. The modal chromosome number of LT-D is 40, and this number is stable during at least 12 mo of continuous culture. LT-D cells are morphologically distinct from embryonal carcinoma cells and no longer express the SSEA-1 cell surface antigen or high alkaline phosphatase activity characteristic of embryonal carcinoma cells. LT-D cells can be induced to fuse into structures resembling myotubes. The formation of these structures is accompanied by expression of the muscle-specific isozyme of creatine phosphokinase and desmin, a muscle-specific component of intermediate filaments. Lastly, LT-D cells do not form tumors in syngenetic mice.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 6","pages":"463-72"},"PeriodicalIF":0.0,"publicationDate":"1984-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02619619","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17213065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Streptomycin retards the phenotypic maturation of chick myogenic cells. 链霉素延缓鸡肌原细胞的表型成熟。
Pub Date : 1984-06-01 DOI: 10.1007/BF02619620
P S Moss, D H Spector, C A Glass, R C Strohman

As part of an effort to optimize conditions required for the complete maturation of muscle cells in vitro, we have investigated the effects of the antibiotics penicillin, streptomycin, and Fungizone (amphotericin B) on the development of cultured chick embryo skeletal muscle. It is shown that even low dosages of streptomycin, but not penicillin or Fungizone, retard protein synthesis and accumulation in these cultures. Myosin accumulation was also reduced and the appearance of striations in fused cells was delayed in myotubes formed in medium containing streptomycin. Additional data suggest that this overall retardation of myogenesis is due to the influence of streptomycin on maturing myotubes rather than early proliferation and cell fusion. These results are discussed with regard to recent efforts to promote the full maturation of muscle cells grown in culture.

为了优化体外肌肉细胞完全成熟所需的条件,我们研究了抗生素青霉素、链霉素和真菌素(两性霉素B)对培养鸡胚胎骨骼肌发育的影响。研究表明,即使是低剂量的链霉素,而不是青霉素或真菌素,也会延缓这些培养物中蛋白质的合成和积累。在含有链霉素的培养基中形成的肌管中,肌球蛋白的积累也减少,融合细胞的条纹出现延迟。另外的数据表明,这种肌肉发生的整体迟缓是由于链霉素对成熟肌管的影响,而不是早期增殖和细胞融合。这些结果讨论了最近的努力,以促进肌肉细胞在培养中生长的完全成熟。
{"title":"Streptomycin retards the phenotypic maturation of chick myogenic cells.","authors":"P S Moss,&nbsp;D H Spector,&nbsp;C A Glass,&nbsp;R C Strohman","doi":"10.1007/BF02619620","DOIUrl":"https://doi.org/10.1007/BF02619620","url":null,"abstract":"<p><p>As part of an effort to optimize conditions required for the complete maturation of muscle cells in vitro, we have investigated the effects of the antibiotics penicillin, streptomycin, and Fungizone (amphotericin B) on the development of cultured chick embryo skeletal muscle. It is shown that even low dosages of streptomycin, but not penicillin or Fungizone, retard protein synthesis and accumulation in these cultures. Myosin accumulation was also reduced and the appearance of striations in fused cells was delayed in myotubes formed in medium containing streptomycin. Additional data suggest that this overall retardation of myogenesis is due to the influence of streptomycin on maturing myotubes rather than early proliferation and cell fusion. These results are discussed with regard to recent efforts to promote the full maturation of muscle cells grown in culture.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 6","pages":"473-8"},"PeriodicalIF":0.0,"publicationDate":"1984-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02619620","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17799154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 10
Characterization of a stable, anchorage-dependent clone obtained from a spontaneously transformed mouse cell line. 从自然转化的小鼠细胞系中获得的稳定的、依赖锚定的克隆的特性。
Pub Date : 1984-06-01 DOI: 10.1007/BF02619621
R Godbout, B L Gallie, R A Phillips

A variant nontransformed clone, I21, was selected from the spontaneously transformed mouse fibroblast line, IT22. Selection was done by plating IT22 in methylcellulose and picking single cells after 2 d. Cultures derived from these single cells were selected again and one clone, I21, derived from the second round of selection was characterized extensively. I21 and IT22 have the same plating efficiency (PE) on plastic, but in agarose they differ by 1000-fold. In comparison to IT22, I21 has a normal morphological appearance, a lower saturation density, a higher viability in stationary phase, an increased doubling time, an increased chromosome content, and is unable to form tumors in nude mice. I21 has remained remarkably stable in culture and has not reverted to the transformed phenotype for at least 300 generations in culture. Over 100 clones of I21, expanded to 10(6) cells, failed to show an increased PE in agarose. Even expansion of the rare colonies of I21 that grow in agarose failed to produce clones similar to IT22.

从自发转化的小鼠成纤维细胞系IT22中选择了一个变异的非转化克隆I21。选择方法是将IT22镀在甲基纤维素中,并在2天后挑选单细胞。从这些单细胞中获得的培养物再次进行选择,从第二轮选择中获得的一个克隆I21进行了广泛的鉴定。I21和IT22在塑料上具有相同的镀效率(PE),但在琼脂糖上却相差1000倍。与IT22相比,I21形态外观正常,饱和密度较低,静止期活力较高,倍增时间增加,染色体含量增加,在裸鼠体内不能形成肿瘤。I21在培养中保持了显著的稳定,并且在培养中至少有300代没有恢复到转化的表型。超过100个I21克隆,扩增到10(6)个细胞,琼脂糖中的PE没有增加。即使对生长在琼脂糖中的I21的罕见菌落进行扩增,也无法产生与IT22相似的克隆。
{"title":"Characterization of a stable, anchorage-dependent clone obtained from a spontaneously transformed mouse cell line.","authors":"R Godbout,&nbsp;B L Gallie,&nbsp;R A Phillips","doi":"10.1007/BF02619621","DOIUrl":"https://doi.org/10.1007/BF02619621","url":null,"abstract":"<p><p>A variant nontransformed clone, I21, was selected from the spontaneously transformed mouse fibroblast line, IT22. Selection was done by plating IT22 in methylcellulose and picking single cells after 2 d. Cultures derived from these single cells were selected again and one clone, I21, derived from the second round of selection was characterized extensively. I21 and IT22 have the same plating efficiency (PE) on plastic, but in agarose they differ by 1000-fold. In comparison to IT22, I21 has a normal morphological appearance, a lower saturation density, a higher viability in stationary phase, an increased doubling time, an increased chromosome content, and is unable to form tumors in nude mice. I21 has remained remarkably stable in culture and has not reverted to the transformed phenotype for at least 300 generations in culture. Over 100 clones of I21, expanded to 10(6) cells, failed to show an increased PE in agarose. Even expansion of the rare colonies of I21 that grow in agarose failed to produce clones similar to IT22.</p>","PeriodicalId":13317,"journal":{"name":"In Vitro","volume":"20 6","pages":"479-85"},"PeriodicalIF":0.0,"publicationDate":"1984-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF02619621","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17799155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
期刊
In Vitro
全部 Acc. Chem. Res. ACS Applied Bio Materials ACS Appl. Electron. Mater. ACS Appl. Energy Mater. ACS Appl. Mater. Interfaces ACS Appl. Nano Mater. ACS Appl. Polym. Mater. ACS BIOMATER-SCI ENG ACS Catal. ACS Cent. Sci. ACS Chem. Biol. ACS Chemical Health & Safety ACS Chem. Neurosci. ACS Comb. Sci. ACS Earth Space Chem. ACS Energy Lett. ACS Infect. Dis. ACS Macro Lett. ACS Mater. Lett. ACS Med. Chem. Lett. ACS Nano ACS Omega ACS Photonics ACS Sens. ACS Sustainable Chem. Eng. ACS Synth. Biol. Anal. Chem. BIOCHEMISTRY-US Bioconjugate Chem. BIOMACROMOLECULES Chem. Res. Toxicol. Chem. Rev. Chem. Mater. CRYST GROWTH DES ENERG FUEL Environ. Sci. Technol. Environ. Sci. Technol. Lett. Eur. J. Inorg. Chem. IND ENG CHEM RES Inorg. Chem. J. Agric. Food. Chem. J. Chem. Eng. Data J. Chem. Educ. J. Chem. Inf. Model. J. Chem. Theory Comput. J. Med. Chem. J. Nat. Prod. J PROTEOME RES J. Am. Chem. Soc. LANGMUIR MACROMOLECULES Mol. Pharmaceutics Nano Lett. Org. Lett. ORG PROCESS RES DEV ORGANOMETALLICS J. Org. Chem. J. Phys. Chem. J. Phys. Chem. A J. Phys. Chem. B J. Phys. Chem. C J. Phys. Chem. Lett. Analyst Anal. Methods Biomater. Sci. Catal. Sci. Technol. Chem. Commun. Chem. Soc. Rev. CHEM EDUC RES PRACT CRYSTENGCOMM Dalton Trans. Energy Environ. Sci. ENVIRON SCI-NANO ENVIRON SCI-PROC IMP ENVIRON SCI-WAT RES Faraday Discuss. Food Funct. Green Chem. Inorg. Chem. Front. Integr. Biol. J. Anal. At. Spectrom. J. Mater. Chem. A J. Mater. Chem. B J. Mater. Chem. C Lab Chip Mater. Chem. Front. Mater. Horiz. MEDCHEMCOMM Metallomics Mol. Biosyst. Mol. Syst. Des. Eng. Nanoscale Nanoscale Horiz. Nat. Prod. Rep. New J. Chem. Org. Biomol. Chem. Org. Chem. Front. PHOTOCH PHOTOBIO SCI PCCP Polym. Chem.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1