2018
DOI: 10.1016/j.gendis.2018.04.003
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Establishment and functional characterization of the reversibly immortalized mouse glomerular podocytes (imPODs)

Abstract: Glomerular podocytes are highly specialized epithelial cells and play an essential role in establishing the selective permeability of the glomerular filtration barrier of kidney. Maintaining the viability and structural integrity of podocytes is critical to the clinical management of glomerular diseases, which requires a thorough understanding of podocyte cell biology. As mature podocytes lose proliferative capacity, a conditionally SV40 mutant tsA58-immortalized mouse podocyte line (designated as tsPC) was es… Show more

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Cited by 23 publications
(30 citation statements)
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“…The use of the retroviral vector SSR #41 to express SV40 T antigen flanked with the FRT sites have been described previously. [15][16][17][18] Briefly, the SSR #41 vector and pCL-Ampho packaging vector were co-transfected into293 Phoenix Ampho (293PA) cells to produce the packaged retrovirus. Exponentially growing TGMCs were infected with the SSR #41 retrovirus and subjected to hygromycin B selection (0.4mg/mL) for 3-5 days twice in complete DMEM at 37°C, yielding the stably immortalized tooth germ mesenchymal cells, designated as the iTGMC line.…”
Section: Establishment Of Reversibly Immortalized Tooth Germ Mesencmentioning
confidence: 99%
“…The use of the retroviral vector SSR #41 to express SV40 T antigen flanked with the FRT sites have been described previously. [15][16][17][18] Briefly, the SSR #41 vector and pCL-Ampho packaging vector were co-transfected into293 Phoenix Ampho (293PA) cells to produce the packaged retrovirus. Exponentially growing TGMCs were infected with the SSR #41 retrovirus and subjected to hygromycin B selection (0.4mg/mL) for 3-5 days twice in complete DMEM at 37°C, yielding the stably immortalized tooth germ mesenchymal cells, designated as the iTGMC line.…”
Section: Establishment Of Reversibly Immortalized Tooth Germ Mesencmentioning
confidence: 99%
“…To assess the duration of adenovirus-mediated gene expression in liver, we injected the CsCl gradient purified Ad-FLuc into mouse liver (4-week-old male, 10 10 pfu/injection/mouse). Whole body optical imaging was performed at day 5 after injection by using D-Luciferin Potassium (Gold Biotechnology Inc.) as luciferase substrate and assessed with the Xenogen IVIS 200 Imaging System as described [6770].…”
Section: Methodsmentioning
confidence: 99%
“… 50 , 51 The above cells were cultured in DMEM containing 10% fetal bovine serum (FBS, Gemini Bio-Products, West Sacramento, CA, USA), supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin at 37°C in 5% CO 2 as described. 19 , 26 , 41 , 52 , 53 , 54 Restriction endonucleases, NEBuilder HiFi DNA assembly master mix, Phusion high-fidelity DNA polymerase, and the electrocompetent DH10B cells were purchased from New England Biolabs (NEB, Ipswich, MA, USA). Blasticidin S was ordered from InvivoGen (San Diego, CA, USA).…”
Section: Methodsmentioning
confidence: 99%
“…The retroviral vector-mediated transduction and establishment of stable cell lines were carried out as previously described. 19 , 22 , 53 , 54 , 57 , 58 , 59 , 60 Briefly, the retroviral transfer vectors pSiEB-simSmad4, pSiEB-simSmad1458, and pSiEB-simSmad4158 were co-transfected with pCL-Ampho into subconfluent 293 Phoenix Ampho (293PA) cells to package retroviruses. The retrovirus-containing supernatants were harvested at 36–72 h after transfection and pooled to infect subconfluent BMSCs.…”
Section: Methodsmentioning
confidence: 99%