Aortic media explants cell proliferation and production of mucopolysaccharides, collagen, and elastic tissue

Jarmolych, J.; Daoud, A.S.; Landau, Joseph V.; Fritz, Katherine; McElvene, E.

doi:10.1016/0014-4800(68)90033-6

Cited by 110 publications

(27 citation statements)

References 7 publications

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“…We therefore designated these changes in marker gene expression as the early phase of PM. Early PM has been implicated in earlier studies in which the authors noticed a small number of phenotypically modulated cells with fewer myofilaments in the 1-day culture of free-floating explants [68,69]. To the best of our knowledge, this finding has not been pursued further.…”

Section: Discussionmentioning

confidence: 90%

The early- and late stages in phenotypic modulation of vascular smooth muscle cells: Differential roles for lysophosphatidic acid

Guo

Makarova

Cheng

et al. 2008

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Lysophosphatidic acid (LPA) has been implicated as causative in phenotypic modulation (PM) of cultured vascular smooth muscle cells (VSMC) in their transition to the dedifferentiated phenotype. We evaluated the contribution of the three major LPA receptors, LPA 1 and LPA 2 GPCR and PPARγ, on PM of VSMC. Expression of differentiated VSMC-specific marker genes, including smooth muscle α-actin, smooth muscle myosin heavy chain, calponin, SM-22α, and hcaldesmon, was measured by quantitative real-time PCR in VSMC cultures and aortic rings kept in serum-free chemically defined medium or serum-or LPA-containing medium using wild-type C57BL/6, LPA 1 , LPA 2 , and LPA 1&2 receptor knockout mice. Within hours after cells were deprived of physiological cues, the expression of VSMC marker genes, regardless of genotype, rapidly decreased. This early PM was neither prevented by IGF-I, inhibitors of p38, ERK1/2, or PPARγ nor significantly accelerated by LPA or serum. To elucidate the mechanism of PM in vivo, carotid artery ligation with/without replacement of blood with Krebs solution was used to evaluate contributions of blood flow and pressure. Early PM in the common carotid was induced by depressurization regardless of the presence/absence of blood, but eliminating blood flow while maintaining blood pressure or after sham surgery elicited no early PM. The present results indicate that LPA, serum, dissociation of VSMC, IGF-I, p38, ERK1/2, LPA 1 , and LPA 2 are not causative factors of early PM of VSMC. Tensile stress generated by blood pressure may be the fundamental signal maintaining the fully differentiated phenotype of VSMC.

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Section: Discussionmentioning

confidence: 90%

The early- and late stages in phenotypic modulation of vascular smooth muscle cells: Differential roles for lysophosphatidic acid

Guo

Makarova

Cheng

et al. 2008

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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“…Culture of these cells has been attempted by a number of investigators (20)(21)(22)(23)(24) over the last few years. but two recurring problems have hampered this work: (a) inability to maintain endothelial cells in pure culture for reasonable periods of time; (b) inability to identify the cultured cells as endothelium.…”

Section: Introductionmentioning

confidence: 99%

Culture of Human Endothelial Cells Derived from Umbilical Veins. IDENTIFICATION BY MORPHOLOGIC AND IMMUNOLOGIC CRITERIA

Jaffe¹,

Nachman²,

Becker³

et al. 1973

J. Clin. Invest.

6,471

2,694

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“…The ability of these cells to synthesize collagen was suggested initially by morphologic observations of collagen fibrils in close association with smooth muscle cells of the media of the arterial wall, an area in which these cells appear to be the sole type present (1)(2)(3). Collagen fibrils were also seen in cultures of smooth muscle cells derived from medial explants of smooth muscle (4). Recent studies have identified three genetically distinct collagens in the arterial wall: type I, composed of two al(I) chains and one a2 chain; type III, consisting of three identical al(III) chains; and type IV collagen, associated with basement membranes (5,6).…”

mentioning

confidence: 95%

Biosynthesis of type I and III collagens by cultured smooth muscle cells from human aorta.

Layman¹,

Epstein²,

Dodson³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

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The synthesis of collagen by human aortic smooth muscle cells was studied after incubating the cells with [3HIproline and [3Hlglycine for 48 hr. The culture medium and cells were lyophilized dthen digested with cyanogen bromide (CNBr) in 70% (wt/vol) formic acid. The resultant peptides were subjected to ion exchange chromatography on CM-cellulose and gel filtration on agarose. On the basis of the molar ratios of the al(Il)CB8 and al(III)CB8 peptides of the al(I) and al (III) chains, approximately one quarter of the collagen synthesized by these cells was identified as type III and three quarters as type I. These data indicate that the smooth muscle can synthesize at least two types of collagen found in the arterial wall. MATERIAL AND METHODS Culture Techniques. Cultures of smooth muscle cells were established from explants of medial smooth muscle obtained from adult human aortas 4-6 hr postmortem. The method for isolating these cells in culture was similar to that of Ross (9), with the exception that the growth medium was supplemented with pooled adult human serum. McCoy's 5a modified medium (Gibco) containing 20% human serum (heat inactivated), 200 units/ml of penicillin, 200 ,g/ml of streptomycin, and 30 units/ml of mycostatin was used for culturing the explanted aortic smooth muscle. Thereafter, 10% human serum was employed for subculturing the smooth muscle cells. Culture medium was changed twice weekly. Only cultures between the third and sixth passage (1:2 split) and exhibiting morphologic 671 and growth characteristics reported for smooth muscle cells were used in these studies.Studies After 48 hr, the medium was removed and lyophilized. The cell layer was scraped from the flask in 20 ml of distilled water and lyophilized. In some experiments both the medium and the cell layer were pooled and lyophilized. The lyophilized samples were digested with CNBr in 70% (wt/vol) formic acid, desalted, and lyophilized as described previously (10). The lyophilized peptides and carrier peptides derived from CNBr digestion of insoluble human placental membranes were redissolved in starting buffer (0.02 M sodium citrate, 0.02 M NaCl adjusted to pH 3.8 with citric acid) and chromatographed on CM-cellulose as previously described (10). Further separation and determination of the relative quantities of CNBr-liberated peptides derived from CM-cellulose chromatography were achieved on a column (1.8 X 230 cm) of agarose (Bio-Gel A-1.5m, 200400 mesh) at room temperature (10). The relative quantities of peptides present were compared by tracing the appropriate peaks onto paper, cutting out the traced outlines, and weighing the individual pieces (10). Some samples were hydrolyzed in 6 M HCI for 24 hr at 1100 and analyzed for radioactive proline and hydroxyproline on a Beckman model 119 amino acid analyzer equipped with a stream-splitting device.Electron Microscopy. For fine structural studies cultured smooth muscle cells were fixed in situ with 3% (vol/vol) glutaraldehyde in 0.1 M phosphate buffer, pH 7.4. The cells w...

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Aortic media explants cell proliferation and production of mucopolysaccharides, collagen, and elastic tissue

Cited by 110 publications

References 7 publications

The early- and late stages in phenotypic modulation of vascular smooth muscle cells: Differential roles for lysophosphatidic acid

The early- and late stages in phenotypic modulation of vascular smooth muscle cells: Differential roles for lysophosphatidic acid

Culture of Human Endothelial Cells Derived from Umbilical Veins. IDENTIFICATION BY MORPHOLOGIC AND IMMUNOLOGIC CRITERIA

Biosynthesis of type I and III collagens by cultured smooth muscle cells from human aorta.

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