Cell Culture Protocols for Fibroblast Models

Normal rat kidney fibroblasts (NRK-49F, ATCC, Manassas, VA, USA) and mouse embryonic fibroblasts (NIH-3T3, ATCC) were grown in Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) containing 5% fetal bovine serum (FBS) and 1% antibiotics [29 (link),30 (link)]. Human embryonic kidney (HEK) 293T (ATCC) cells were cultured in DMEM with 10% FBS and 1% antibiotics. The cells were maintained in an incubator at 37 °C with 5% CO₂. The NRK-49F and NIH-3T3 cells were incubated overnight in starvation media containing DMEM and 0.1% FBS before TGF-β1 (Peprotech, Rocky Hill, NJ, USA) treatment.

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Wen J., Jiao B., Tran M, & Wang Y. (2022). Pharmacological Inhibition of S100A4 Attenuates Fibroblast Activation and Renal Fibrosis. Cells, 11(17), 2762.

Publication 2022

NRK-49F NIH-3T3 DMEM HEK) 293T TGF-β1

1 fetal Antibiotics Cells Eagle Embryonic Fetal bovine serum Fibroblasts Human Kidney Mouse Nih 3t3 cells

Corresponding Organization : VA Connecticut Healthcare System

Other organizations : Guangdong Academy of Medical Sciences, Guangdong Provincial People's Hospital

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Variable analysis

independent variables

TGF-β1 treatment

dependent variables

Not explicitly mentioned

control variables

Cell type (NRK-49F, NIH-3T3, HEK 293T)
Cell culture medium (DMEM with 5% or 10% FBS and 1% antibiotics)
Cell culture conditions (37 °C, 5% CO2)
Starvation media (DMEM with 0.1% FBS) for NRK-49F and NIH-3T3 cells prior to TGF-β1 treatment

controls

Positive control: Not specified
Negative control: Not specified

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