Mesangial cell behavior in a three-dimensional extracellular matrix

Kitamura, Masanori; Mitarai, Tetsuya; Maruyama, Naoki; Nagasawa, Ryo; Yoshida, Hiroaki; Sakai, Osamu

doi:10.1038/ki.1991.257

Cited by 58 publications

(29 citation statements)

References 24 publications

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“…Rat mesangial cells were established from isolated renal glomeruli of an adult male Sprague-Dawley rat (7). A cell clone SM43 was established by a limiting dilutional method and identified as being of mesangial cell phenotype as described (25). By using a modified calcium phosphate coprecipitation method (26), SM43 mesangial cells were cotransfected with a regulator plasmid pUHD15-1 encoding tTA (21) and a plasmid pRc/CMV (Invitrogen) that introduces a neomycin phosphotransferase gene.…”

Section: Methodsmentioning

confidence: 99%

Creation of a reversible on/off system for site-specific in vivo control of exogenous gene activity in the renal glomerulus.

Kitamura

1996

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

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Using genetically engineered glomerular mesangial cells, an in vivo gene transfer approach was developed that specifically targets the renal glomerulus. By combining this system with a tetracycline (Tc)-responsive promoter, the present study aimed to create a reversible on/off system for site-specific in vivo control of exogenous gene activity within the glomerulus. In the Tc regulatory system, a Tc-controlled transactivator (tTA) encoded by a regulator plasmid induces target gene transcription by binding to a tTA-responsive promoter located in a response plasmid. Tc inhibits this tTA-dependent transactivation via its affilnity for tTA. In double-transfected cells, therefore, the activity of a transgene can be controlled by Tc. Cultured rat mesangial cells were cotransfected with a regulator plasmid and a response plasmid that introduces a 18-galactosidase gene. In vitro, stable double-transfectant MtTAG cells exhibited no .8-galactosidase activity in the presence of Tc. However, following withdrawal of Tc from culture media, expression of f8-galactosidase was induced within 24 h. When Tc was again added, the expression was rapidly resuppressed. Low concentrations of Tc were sufficient to maintain the silent state of tTA-dependent promoter. MtTAG cells were then transferred into the rat glomeruli via renal artery injection. In the isolated chimeric glomeruli, expression of 8-galactosidase was induced ex vivo in the absence of Tc, whereas it was repressed in its presence. When Tc-pretreated MtTAG cells were transferred into the glomeruli of untreated rats, 18-galactosidase expression was induced in vivo within 3 days. Oral administration of Tc dramatically suppressed this induction. These data demonstrate the feasibility of using mesangial cell vectors combined with the Tc regulatory system for site-specific in vivo control of exogenous gene expression in the glomerulus.The renal glomerulus is a microscopic vascular structure scattered throughout the cortex of the kidney. During the past decade, numerous studies have been done to elucidate molecular mechanisms of glomerular diseases. Among various approaches utilized, in vivo gene transfer technology would be one of the most powerful and promising strategies, since this allows us to modulate activity of a certain molecule within the glomerulus (1, 2). However, recent investigations have encountered some hurdles. Because of the microscopic size (100 jam in diameter) and number (3 x 104 to 1 X 106/kidney) of glomeruli, it is difficult to efficiently and site-specifically deliver foreign genes into the majority of the glomeruli. Conventional cationic liposomes or adenoviral vectors, the most popular in vivo gene transfer vectors, were found to be inefficient for this purpose (3,4). A modified liposome system could be useful (5), but exogenous gene expression has been limited to several days (6). To overcome these problems, I established a novel gene transfer system that specifically targets the glomerulus (7-9). In this method, the glomerular mesangial cell ...

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Section: Methodsmentioning

confidence: 99%

Creation of a reversible on/off system for site-specific in vivo control of exogenous gene activity in the renal glomerulus.

Kitamura

1996

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

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“…Rat mesangial cells were established from isolated renal glomeruli of an adult male Sprague-Dawley rat (27). A cell clone SM43 was established by a limiting dilution method (1) and identified as being of mesangial cell phenotype as described (27). Using a modified calcium phosphate coprecipitation method (28), SM43 mesangial cells were transfected with an expression plasmid pSRE-lacZ (29).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Creation of an In vivo cytosensor using engineered mesangial cells. Automatic sensing of glomerular inflammation controls transgene activity.

Kitamura¹,

Kawachi²

1997

J. Clin. Invest.

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“…In the particular case of mesangial cells, different phenotypic changes have been described when cells were grown under these conditions [10, 11, 12, 13]. For instance, Saito et al [11]demonstrated that the mesangial cell proliferation is different when collagen type I or collagen type III, both are often observed in diseased glomeruli, is present.…”

Section: Introductionmentioning

confidence: 99%

Phenotypic Modifications of Human Mesangial Cells by Extracellular Matrix: The Importance of Matrix in the Contractile Response to Reactive Oxygen Species

Cruz

Ruiz-Torres²,

Lucio-Cazaña³

et al. 2000

Nephron Exp Nephrol

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The progression of chronic renal diseases is characterized by the accumulation of extracellular matrix proteins in the glomerulus. The present experiments were designed to analyze the effect of hydrogen peroxide on the contractile and proliferative phenotypes of human mesangial cells grown on different culture substrates: plastic, collagen type I, and collagen type IV. Contraction was analyzed by measuring planar cell surface area and myosin light chain phosphorylation, whereas proliferation was studied by [³H]thymidine incorporation. No changes were detected in the proliferation rate of human mesangial cells grown on different culture substrates, neither under basal conditions nor in the presence of fetal calf serum or H₂O₂. Cells grown on plastic or collagen did not contract in the presence of H₂O₂, but cells grown on collagen I elicited a significant contraction with H₂O₂. Platelet-activating factor induced contraction of human mesangial cells on the three culture substrates. The different contractile responses observed were not due to different degradation rates of H₂O₂. The present experiments support the importance of extracellular matrix in the response to exogenous stimuli and point to the possibility that patients with changes in the mesangial matrix as a result of chronic renal diseases may have an increased susceptibility to the pathological actions of reactive oxygen species.

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Mesangial cell behavior in a three-dimensional extracellular matrix

Cited by 58 publications

References 24 publications

Creation of a reversible on/off system for site-specific in vivo control of exogenous gene activity in the renal glomerulus.

Creation of a reversible on/off system for site-specific in vivo control of exogenous gene activity in the renal glomerulus.

Creation of an In vivo cytosensor using engineered mesangial cells. Automatic sensing of glomerular inflammation controls transgene activity.

Phenotypic Modifications of Human Mesangial Cells by Extracellular Matrix: The Importance of Matrix in the Contractile Response to Reactive Oxygen Species

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