共查询到20条相似文献,搜索用时 15 毫秒
1.
Keith L. Hall Joseph W. Harding Howard L. Hosick 《In vitro cellular & developmental biology. Animal》1991,27(10):791-798
Summary Vascular smooth muscle cells were isolated from the aortas of spontaneously hypertensive rats and normotensive Wistar-Kyoto
rats by use of the explant method on collagen gels. Clonal cell lines derived from these enriched populations possessed ultrastructural
characteristics of vascular smooth muscle cells in culture; they grew in hill and valley configuration, immunostained with
the muscle actin antibody HHF35, and failed to react with von Willebrand Factor VIII antibody. Fourteen clonal cell lines
were characterized for growth and ligand binding characteristics. Large variations in growth rate and cell density at saturation
were exhibited by clones of both strains. Similar variability was noted for specific binding of endothelial 1 and Sar1,Ile8-angiotensin II to their receptors, indicating considerable phenotypic heterogeneity among the clonal cell lines. Six selected
clones were further characterized for angiotensin II receptor linkage to G proteins. Cells of both strains exhibited comparable
affinity shifts in the presence of GTPγS. These clonal cell lines should be useful for a variety of analyses of the comparative
biology of aortic cells. It is possible that the diversity of phenotypic traits exhibited by these clones reflects the heterogeneity
of vascular smooth muscle tissue found in vivo. 相似文献
2.
E. Jane Ormerod Philip S. Rudland 《In vitro cellular & developmental biology. Plant》1985,21(3):143-153
Summary Single-cell-cloned cell lines have been established from primary cultures of neonatal rat mammary glands. A representative
cuboidal cell line, Rama 704, shows the presence of intermediate filamental proteins keratin and vimentin, and occasional
cells express milk fat globule membrane antigens on their apical surfaces. Rama 704 cells grow as a cuboidal pavement in culture
and produce hemispherical blisters or domes when confluent. Noteworthy ultrastructural features are the presence of junctional
complexes, desmosomes, and apical microvilli typical of epithelia. Cells seeded within floating collagen gels with form a
variety of multicellular outgrowths, some of which are ductlike in morphology and are composed of polarized cells surrounding
a central lumen. The cuboidal cells produce elongated cells under conditions of high cell density and also when cells float
off collagen gels and reattach to the plastic substrate. The former elongated cells have been cloned and three cel lines established:
Rama 710, 711, and 712; the latter uncloned elongated cells are termed Rama 704E. The cloned elongated cells show an increase
in the amounts of basement membrane proteins deposited, a lack of junctional complexes and microvilli, and an increase in
the amount of rough endoplasmic reticulum compared with their parental cells. Rama 704E cells show an enhanced deposition
of basement membrane proteins and increased amounts of actin in the cytoplasm over the elongated cell lines and contain microfilaments
and pincocytotic vesicles similar to those seen in myoepithelial cells. All the elongated cells and lines fail to form ductlike
structures within collagen gels. None of the cell lines form tumors in syngeneic rats although they all produce some tumors
in nude mice, which are composed of cords of epithelioid cells and spindle cells in varying proportions. In addition, some
of the Rama 704 tumors contain rhabdomyoblastic elements that penetrate the host fat pad. This is the first report of the
isolation and characterization of a stable cuboidal cell line from a neonatal rat mammary gland. The Rama 704 cell line shows
morphological and biochemical features of mammary epithelial cells and converts at high cell density to elongated cells that
have also been cloned. 相似文献
3.
4.
Bruno Costa-Silva Meline Coelho da Costa Fernanda Rosene Melo Marcio Alvarez-Silva Giordano Wosgrau Calloni Andréa Gonçalves Trentin 《Experimental cell research》2009,315(6):955-967
The neural crest (NC) is a model system used to investigate multipotency during vertebrate development. Environmental factors control NC cell fate decisions. Despite the well-known influence of extracellular matrix molecules in NC cell migration, the issue of whether they also influence NC cell differentiation has not been addressed at the single cell level. By analyzing mass and clonal cultures of mouse cephalic and quail trunk NC cells, we show for the first time that fibronectin (FN) promotes differentiation into the smooth muscle cell phenotype without affecting differentiation into glia, neurons, and melanocytes. Time course analysis indicated that the FN-induced effect was not related to massive cell death or proliferation of smooth muscle cells. Finally, by comparing clonal cultures of quail trunk NC cells grown on FN and collagen type IV (CLIV), we found that FN strongly increased both NC cell survival and the proportion of unipotent and oligopotent NC progenitors endowed with smooth muscle potential. In contrast, melanocytic progenitors were prominent in clonogenic NC cells grown on CLIV. Taken together, these results show that FN promotes NC cell differentiation along the smooth muscle lineage, and therefore plays an important role in fate decisions of NC progenitor cells. 相似文献
5.
大鼠细小肺动脉平滑肌细胞原代培养和鉴定方法的研究 总被引:2,自引:0,他引:2
目的:建立一种重复性好、培养周期短及传代次数多的大鼠细小肺动脉平滑肌细胞(PASMCs)培养方法。方法:在无菌条件下,分离雄性SD大鼠肺细小动脉,剥离外膜和剔除内皮细胞,经胶原酶I消化,培养PASMCs。0.4%台盼蓝染色测定细胞活力;倒置相差显微镜观察;免疫细胞化学法和免疫荧光染色法,进行平滑肌α-肌动蛋白(α-SMactin)鉴定。结果:形态学观察、免疫细胞化学法及免疫荧光染色法鉴定表明培养细胞为PASMCs;细胞存活率在96.5%以上;原代培养后4~7d即可传代,并且生长特点、细胞形态不易发生改变。结论:采用胶原酶I消化法培养PASMCs,方法简单、酶消化时间易控制、培养周期短、重复性好,培养的原代PASMCs具有数量多和生长迅速的特点。 相似文献
6.
Effects of collagen gel configuration on behavior of vascular smooth muscle cells in vitro: Association with vascular morphogenesis 总被引:5,自引:0,他引:5
Song J Rolfe BE Hayward IP Campbell GR Campbell JH 《In vitro cellular & developmental biology. Animal》2000,36(9):600-610
Summary The growth, behavior, and contractile protein expression of rabbit aortic smooth muscle cells (SMC) grown on, between layers,
or within a collagen gel was investigated by confocal laser scanning fluorescence microscopy and Western analysis. SMC grown
on collagen gel behaved similarly to those on conventional culture dishes. However, when a second layer of collagen was overlaid,
cells underwent an elongated quiescent phase before onset of proliferation and a more than threefold lower logarithmic growth
rate was observed. These cells self-organized into a network with ring-like structures. With increasing culture time, some
of the rings developed into funnel-like, incomplete or complete tubular structures. If a tubular template preexisted within
the gel, the SMC established a cylinder-shaped tube with several circularly arranged muscular layers (similar to an artery
wall). This behavior mimicked endothelial cells during angiogenesis in vitro. A similar phenomenon occurred in cultures in
which SMC were randomly mixed in a collagen gel, but here their behavior and morphology varied with their position within
the gel. Western blot analysis showed that the SMC differentiation marker, smooth muscle myosin heavy chain-2 (SM-2), rapidly
decreased, disappearing by day 10 in SMC grown on collagen, but was still detectable until day 25 in cells cultured between
or within the same gel. These findings indicate that like endothelial cells, vascular SMC can display blood vessel formation
behavior in vitro when an appropriate three-dimensional matrix environment is provided to keep them in a relatively higher-differentiated
and low-proliferative state. 相似文献
7.
8.
Steven A. Moore Arthur R. Strauch Elizabeth J. Yoder Peter A. Rubenstein Michael N. Hart 《In vitro cellular & developmental biology. Plant》1984,20(6):512-520
Summary Cerebral endothelium is being studied rather extensively in tissue culture, but no reports are available describing the tissue
culture of cerebral microvascular smooth muscle. The present paper describes for the first time the isolation and culture
of non-neoplastic mouse cerebral vascular smooth muscle. Microvessels from a dounce homogenate of mouse brain are plated onto
plastic culture dishes in Dulbecco’s modified Eagle media plus 20% fetal bovine serum and treated briefly with collagenase.
Cells migrate from vessels and proliferate sufficiently to be transferred out of primary culture in 2 to 3 wk. Light microscopy
reveals generally broad, polygonal cells that grow collectively in a “hill and valley” pattern. By transmission electron microscopy
the cells possess many characteristics of smooth muscle: basal laminas, clusters of pinocytotic vesicles, and bundles of thin
filaments. Several ill-defined cell-to-cell junctions are also present. Isoelectric focusing and sodium dodecyl sulfate-electrophoresis
of cellular proteins on polyacrylamide gels after pulse labeling cultures with [S-35]methionine demonstrate that these cells actively synthesize a smooth-muscle-specific isoactin, α-actin. The identity of α-actin
is confirmed by analysis of NH2-terminal peptides after actin digestion with trypsin and subsequent peptide cleavage with thermolysin. Both their morphology
and active synthesis of α-actin strongly suggest that these cells are of smooth-muscle origin. Future studies of their metabolism
and interactions with endothelium and astrocytes should provide a better understanding of the cerebral microcirculation.
This work was supported by Veteran’s Administration Research Funds, National Institutes of Health Grant HL 14230 (Arteriosclerosis
Specialized Center of Research, sponsored by the National Heart, Lung, and Blood Institute), and Cardiovascular Research Program
Project Grant from the National Institutes of Health to the University of Iowa (HL-14388). A. R. S. is a postdoctoral fellow
of the National Institute of General Medical Sciences (GM-09020). P. A. R. is an established investigator of the American
Heart Association. 相似文献
9.
Nelly Blaes Marie-Claude Bourdillon Jean-Marie Daniel-Lamaziere Jean-Jacques Michaille Mauricio Andujar Chantal Covacho 《In vitro cellular & developmental biology. Animal》1991,27(9):725-734
Summary Smooth muscle cell proliferation is an important feature of atherogenesis. Some works have hypothesized that a transformation
of smooth muscle cells could arise during this pathological process. The present paper describes two spontaneously transformed
cell lines of arterial smooth muscle cells (SMC) established from aortic media of adult rat. The cell lines have been designated
V6 and V8; some of their morphologic, growth, and metabolic characteristics are described and compared to their parent cells.
The two cell lines appeared distinct by their morphology and by their degree of transformation. V6 cells appeared as elongated
spindle-shaped cells whereas V8 cells were spread cells with a cobblestone pattern. Karyotypes of both cell lines showed a
high polyploidy level. V6 and V8 cell lines were immortalized and showed growth characteristics of transformed cells: low
requirement of serum to grow, ability to form colonies in soft agar and tumorigenicity in nude mice; V8 cells presented a
higher malignancy than V6 cells. Both V6 and V8 cells exhibited characteristics of cultured arterial SMC: ultrastructure,
alpha actin expression at the protein and mRNA level, prostacyclin production. The remarkably different morphologies of the
V6 and V8 lines and their transformed phenotype suggest that these cell lines could be useful models to study SMC differentiation
and proliferation with respect to atherosclerotic or hypertensive vascular diseases.
Electron microscopy was performed in the Center of Electron Microscopy Applied to Biology and Geology (CEMABG), Claude Bernard
University, Lyon I. Flow cytofluorometry was performed in the Center of Fluorometry, Department of Human Biology, Claude Bernard
University, Lyon I and funded by ARC No 6055-80.
This work was supported by INSERM, by MRT grant 86-C-0301 and by ARC grant 415-87. 相似文献
10.
目的:研究胚胎血管发育早期SMα-actin、SM22α、myocardin、平滑肌肌球蛋白重链(SMMHC)的表达规律,并初步探讨在此阶段血小板源性生长因子-BB(PDGF-BB)对血管平滑肌细胞(VSMCs)分化的影响。方法:采用转染平滑肌特异性蛋白SM22α启动子控制下表达增强型绿色荧光蛋白(GFP)报告基因载体的胚胎干细胞制备拟胚体(EBs),用免疫荧光染色、RT-PCR、Western blot分析SMα-actin、SM22α、myocardin、SMMHC的表达时相;然后分别用0μmol/L(对照组)、10μmol/L、50μmol/L AG1296(血小板源性生长因子受体抑制剂)处理EBs,观察三组SMα-actin、SM22α、myocardin、SMMHC在基因及蛋白水平上的表达变化。结果:胚胎血管发育早期SMα-actin、myocardin、SM22α、SMMHC分别在EBs第0(胚胎干细胞)、8、11、13d开始有表达。AG1296三种浓度处理后SMα-actin、myocardin、SM22α、SMMHC蛋白表达及myocardin、SM22α和SMMHC mRNA表达均无明显差异。结论:EBs发育过程中存在着自发的VSMCs分化,SMα-actin表达最早,依次为myocardin、SM22α、SMMHC;PDGF-BB对EBs分化早期VSMCs标志物表达的调控可能不是必要的。 相似文献
11.
Maintenance and characterization of human myometrial smooth muscle cells in monolayer culture 总被引:4,自引:0,他引:4
M. Linette Casey Paul C. MacDonald Murray D. Mitchell Jeanne M. Snyder 《In vitro cellular & developmental biology. Plant》1984,20(5):396-403
Summary Human myometrial cells were dispersed from uterine tissue by limited enzymatic digestion of myometrium that was obtained at
the time of hysterectomy. The dispersed myometrial cells that are obtained in this manner can be maintained in monolayer culture
in the presence of medium that contains fetal bovine serum. In primary culture, as well as after passage, the characteristics
of these cells are morphologically and biochemically similar to those of smooth muscle cells and myometrial tissue.
This investigation was supported, in part, by U.S. Public Health Service Grants 5-P50-HD11149 and 5-P01-AG00306. 相似文献
12.
Rolf Bjerkvig Sverre K. Steinsvåg Ole Didrik Laerum 《In vitro cellular & developmental biology. Plant》1986,22(4):180-192
Summary A stationary tissue culture system for reaggregation cultures of rat brain cells is described. Aggregates were formed by placing
cells at high concentrations in liquid overlay cultures on a nonadherent nutrient agar surface. No physical stress in the
form of rotation or shaking was applied to the aggregating cell population.
Transmission electron microscopy and immunohistochemistry showed that the cells developed from homogeneously dispersed, immature
cells in Day 4 aggregates, to mature astrocytes, oligodendrocytes, and neurons in Day 20 aggregates. Twenty days and older
aggregates had a tightly packed neuropil which was most prominent in a cell-sparse outer layer of the aggregates. When the
aggregates were allowed to adhere to a substrate, both glial fibrillary acidic protein (GFAP) positive and negative cells
were observed migrating out from the aggregates. Cells giving a positive reaction for neuron specific enolase (NSE) were also
present. This reaggregation procedure, with transfer of selected brain cell aggregates into agar-coated multiwells is an alternative
three-dimensional culture system which can be potentially useful in the study of morphogenesis and cell interactions in the
nervous system.
This project was supported by the Norwegian Cancer Society. 相似文献
13.
Paul G. McGuire Helen M. Walker-Caprioglio Sally A. Little Linda J. McGuffee 《In vitro cellular & developmental biology. Animal》1993,29(2):135-139
Summary The structure and function of vascular smooth muscle cells have been extensively investigated with the aid of in vitro culture
techniques. The majority of studies have utilized aortic tissue as the source of cells. We present here a method for isolating
and culturing smooth muscle cells of the rat superior mesenteric artery, an elasto-muscular vessel that is structurally and
functionally different from the aorta. Cells were isolated from partially digested explants and characterized by immunochemical
and biochemical techniques. Unlike cultured fibroblasts, the cultured cells stained positive for smooth muscle specific actin.
The cells also produced laminin and type IV collagen in culture. This method provides a means for the isolation of large numbers
of viable smooth muscle cells from the superior mesenteric artery which can be propagated in culture for in vitro study. 相似文献
14.
Pintucci G Yu PJ Saponara F Kadian-Dodov DL Galloway AC Mignatti P 《Journal of cellular biochemistry》2005,95(6):1292-1300
Basic fibroblast growth factor (FGF-2) and platelet-derived growth factor (PDGF) are implicated in vascular remodeling secondary to injury. Both growth factors control vascular endothelial and smooth muscle cell proliferation, migration, and survival through overlapping intracellular signaling pathways. In vascular smooth muscle cells PDGF-BB induces FGF-2 expression. However, the effect of PDGF on the different forms of FGF-2 has not been elucidated. Here, we report that treatment of vascular aortic smooth muscle cells with PDGF-BB rapidly induces expression of 20.5 and 21 kDa, high molecular weight (HMW) FGF-2 that accumulates in the nucleus and nucleolus. Conversely, PDGF treatment has little or no effect on 18 kDa, low-molecular weight FGF-2 expression. PDGF-BB-induced upregulation of HMW FGF-2 expression is controlled by sustained activation of extracellular signal-regulated kinase (ERK)-1/2 and is abolished by actinomycin D. These data describe a novel interaction between PDGF-BB and FGF-2, and indicate that the nuclear forms of FGF-2 may mediate the effect of PDGF activity on vascular smooth muscle cells. 相似文献
15.
Cartilage tissue differentiation from mesenchymal cells derived from mature muscle in tissue culture 总被引:2,自引:0,他引:2
Marshall R. Urist Yoji Terashima M. Nakagawa Charles Stamos 《In vitro cellular & developmental biology. Plant》1978,14(8):697-706
Summary Under the influence of biochemical components of bone matrix gelatin (BMG), cartilage differentiates in tissue culture from
the connective tissue cell outgrowths of mature muscle. Proliferation and differentiation begin within 24 hr with synthesis
of hyaluronate, continue with high levels of synthesis of DNA and hyaluronidase, and culminate in production of large quantities
of chondroitin sulfate. The addition of hyaluronic acid to the culture medium during the first 48 hr of culture depresses,
whereas chondroitin sulfate enhances, subsequent production of cartilage. These observations on the cell biosynthetic products
prior to the appearance of mature cartilage suggest that the BMG-modified connective tissue outgrowths of mature muscle exhibit
the developmental potential of embryonic axial mesenchyme. Whether muscle harbors embryonic cells in a programmed but not
yet activated readiness (protodifferentiated state) to differentiate into cartilage, or simply contributes a population of
temporarily dedifferentiated fibroblasts, is not known, but in any event, BMG switches the pathway of further development
from fibrous connective tissue to cartilage.
These investigations were supported by grants-in-aid from the USPHS, National Institute of Dental Research (DE-2103-01). Drs.
Terashima and Nakagawa received a research fellowship from the Solo Cup Corporation. Charles Stamos was a Eugene and Marion
Bailey Summer Student Research Fellow. 相似文献
16.
Embryonic data and ultrastructural analyses suggest that the primitive endothelium signals undifferentiated mesenchymal cells to migrate to the forming blood vessel and subsequently regulates mural cell growth and behavior. Upon maturation of the blood vessel, chemotactic and mitogenic signals are apparently diminished and differentiated smooth muscle cells normally remain quiescent. This homeostasis is seemingly upset in conditions which lead to pathologies characterized by smooth muscle cell hyperplasia such as atherosclerosis. By culturing endothelial cells at different densities, we attempted to re-create the various stages of vascular development. Whereas media conditioned by sparse endothelial cells stimulate smooth muscle cells, media conditioned by dense endothelial cell cultures are inhibitory. Culture of sparse smooth muscle cells in media conditioned for 3 days by postconfluent endothelial cell cultures leads to dose-dependent and reversible smooth muscle cell inhibition. Furthermore, in the presence of the endothelial cell-derived inhibitor, smooth muscle cells are rendered refractory to mitogens such as fibroblast growth factor and platelet-derived growth factor. The inhibitory activity is not attributable to the well-characterized inhibitors of smooth muscle cell growth, transforming growth factor type-β, prostaglandin I2, or heparan sulfate proteoglycan. Partial characterization of the inhibitory conditioned media suggests that the active molecule is smaller than 1,000 da, and stable to boiling as well as proteinase K and heparinase digestion. These findings support the concept that there is intercellular communication between endothelial cells and smooth muscle cells and provide evidence for a novel endothelial cell-derived smooth muscle cell growth inhibitor. 相似文献
17.
Novel techniques to establish new insect cell lines 总被引:3,自引:0,他引:3
Lynn DE 《In vitro cellular & developmental biology. Animal》2001,37(6):319-321
Summary The success of insect cell culture is demonstrated by reports of over 500 established cell lines. While established procedures
that can be used for developing new cell lines exist, these usually require some fine-tuning for various tissue sources. This
paper attempts to depict some of the variations that can be applied. 相似文献
18.
Background
We performed a comparative analysis of the genome-wide DNA methylation profiles from three human embryonic stem cell (HESC) lines. It had previously been shown that HESC lines had significantly higher non-CG methylation than differentiated cells, and we therefore asked whether these sites were conserved across cell lines. 相似文献19.
Demetrios P. Matthopoulos Margaret Tzaphlidou Ioannis Leontiou 《Cell biology international》1995,19(6):499-506
Lithium is being used for the treatment of mental diseases and for the attenuation of muelosuppression during chemotherapy. As during long term lithium treatment kidney damage has been reported, we studied morphological alterations in cells of kidney origin after exposure to lithium chloride. Above the level of 4 mmol, lithium has fatal effects in CV1 cells while HeLa cells that are not originating from kidneys, tolerate higher lithium concentrations. Cellular morphology alters during treatment duration. At early stages, cells become flatter on their substrate and upon longer than 4 days treatment begin to detach from their substrate and eventually cell death comes in a concentration dependent manner. The only morphological alteration observed in a lymphoblastoid cell line was a statistically significant cellular swelling. 相似文献
20.
The mitogenic effects of vasoactive neuropeptides on cultured smooth muscle cell lines 总被引:1,自引:0,他引:1
In order to investigate the relationship between the biochemical pathways that characterize contraction and cell growth, we have studied both contraction, mitogenesis and protein synthesis induced by the vasoactive neuropeptides, substance P (SP), calcitonin gene related peptide (CGRP) and vasoactive intestinal polypeptide (VIP) on four different established vascular and nonvascular smooth muscle cell lines. Contraction in vitro was evaluated by light microscopy and recorded photographically. Mitogenesis and protein synthesis were evaluated by [3H]-thymidine incorporation into cells and [3H]-amino acid incorporation into trichloroacetic acid precipitated materials, respectively. SP stimulated mitogenesis of A7r5 cells (embryonic rat aorta), but failed to induce significant contraction of these cells, whereas, SP induced contraction of cultured adult rat vascular smooth muscle cells (VSMC), but failed to stimulate mitogenesis. CGRP and VIP stimulated mitogenesis and protein synthesis, respectively, of DDT1MF-2 cells (hamster vas deferens), but neither induced contraction of this cell line. All three neuropeptides showed no effect on BC3H1 (mouse smooth muscle-like) cells. These results suggest that neuropeptides with vasoactive properties modulate different stages of cellular mitogenic responses which may be regulated by the degree of maturation of smooth muscle cell. 相似文献