renal epithelial cell growth media  (PromoCell)

Journal: bioRxiv

Article Title: Development of high-affinity, single-domain protein binders for neutralizing household allergens

doi: 10.1101/2025.08.03.668213

Figure Lengend Snippet: (a) Thermostability profile of AVA-1-1-C3 as a representative protein, as determined by differential scanning fluorimetry. (b-g) Cytotoxicity of primary human cells following 48-hour exposure to AVA-1-1-C3 or an equivalent concentration of bovine serum albumin (BSA), evaluated using the CellTiter-Glo® (CTG) luminescent cell viability assay. Cell types included human epithelial keratinocytes (HEK, b), human dermal fibroblasts (HDF, c), human follicle dermal papilla cells (HFDPC, d), human renal cortical epithelial cells (HRCEPC, e), human skeletal muscle cells (HSKMC, f), and human dermal microvascular endothelial cells (HDMEC, g). No significant differences in cell viability were observed. Data in (b-g) represents mean ± s.d. and are representative of two experimental replicates.

Article Snippet: Cells were cultured in their corresponding complete media including HDF growth medium (Cell Applications, Cat# 116-500), human EpiVita serum-free growth medium (Cell Applications, Cat# 141-500a), microvascular endothelial cell growth medium (PromoCells, Cat# C-22120), HSkMC growth medium (Cell Applications, Cat# 151-500), and renal epithelial cell growth media (PromoCell, Cat# C-26130).

Techniques: Concentration Assay, Cell Viability Assay

Journal: JCI insight

Article Title: Glycolytic lactate in diabetic kidney disease.

doi: 10.1172/jci.insight.168825

Figure Lengend Snippet: Figure 5. Lactate inhibits mitochondrial function in a dose-dependent manner in human kidney proximal tubule epithelial cells (HK2). (A) Oxygen consumption rates (OCR) were measured in HK2 cells (n = 15) using seahorse extra- cellular flux analyzer. Following basal respiratory measurements, indicated concentrations of lactic acid were injected through injection ports. (B) Basal respiration is represented immediately after treatment with lactate (n = 20). P values in A were calculated using 1-way ANOVA and Dunnett’s test for multiple comparison testing. Data represent mean ± SD. Oligo, oligomycin; FCCP, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone; Rot+AA, Rotenone+Antimycin A.

Article Snippet: PBS-washed sections were transferred into 24-well plate and incubated at 37°C and 5% CO2 with 500 μL of renal epithelial cell growth media (ATCC, PCS-400-040) with NG (7.2 mM) or HG (25 mM) or MCT inhibitor (MilliporeSigma, SR13800) as indicated.

Techniques: Injection, Comparison

Journal: Cell communication and signaling : CCS

Article Title: Connexin-43 hemichannels orchestrate NOD-like receptor protein-3 (NLRP3) inflammasome activation and sterile inflammation in tubular injury.

doi: 10.1186/s12964-023-01245-7

Figure Lengend Snippet: Fig. 5 Peptide5 blocks Cx43-hemichannels to inhibit priming and activation of the NLRP3 inflammasome in primary tubule epithelial cells. Primary hPTECs were treated with pro-fibrotic cytokine TGF-β1 (10 ng/ml) for 48 h in the presence/absence of Peptide5 (25 µM) (a). qRT-PCR analysis of IL-1β, NLRP3 and IL18 mRNA (b) determined a role for Cx43 hemichannel activity in regulation of priming (step 1). Western blot analysis (c-e), IL1β secretion (f) and caspase 1 activity (g) determined an upstream role for Cx43-hemichannel activity in NLRP3 inflammasome activation. These data corroborate our in vivo observations and further suggest that aberrant Cx43 hemichannel activity sits upstream of NRLP3 inflammasome activation. All groups are n = 3–4. An ANOVA and Tukeys post-test was used for all analysis. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001

Article Snippet: Primary hPTECs were maintained in Renal Epithelial Cell Basal Media from ATCC, supplemented with Renal Epithelial Cell Growth Kit components (PCS-400-040) in a humidified atmosphere at 37 °C with 5% CO2.

Techniques: Activation Assay, Quantitative RT-PCR, Activity Assay, Western Blot, In Vivo

Journal: Cell communication and signaling : CCS

Article Title: Connexin-43 hemichannels orchestrate NOD-like receptor protein-3 (NLRP3) inflammasome activation and sterile inflammation in tubular injury.

doi: 10.1186/s12964-023-01245-7

Figure Lengend Snippet: Fig. 8 Peptide5 blocks Cx43-hemichannels in primary hPTECs to dampen secretion of the pro-inflammatory secretome. Primary hPTECs were treated with pro-fibrotic cytokine TGF-β1 (10 ng/ml) for 48 h in the presence/absence of Peptide5 (25µM) (a) and a proteome profiler inflammation array was used to assess secretion of 105 inflammatory mediators (b). Heat map analysis identified multiple changes in protein expression in cells pre-incubated with hemichannel blocker Peptide5 (b), with statistical analysis on 12 selected proteins (c-f) performed. n = 4 per group. Extrapolation of this array data using a published transcriptomic dataset on Nephroseq (g) shows that granulocyte-macrophage colony-stimulating factor (GM-CSF), epithelial cell-derived neutrophil-activating peptide (ENA-78), interleukin-24 (IL24), resistin and leukemia inhibitory factor (LIF) expression are increased in kidneys of patients with CKD (n = 48 in patients with CKD and n = 5 normal controls). All groups are n = 3–4 unless otherwise specified. ANOVA and Tukeys post-test was used for all proteome array analysis (c-f), whilst an unpaired t-test with Welch’s correction was used for analysis of transcriptomic data (g). *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001

Article Snippet: Primary hPTECs were maintained in Renal Epithelial Cell Basal Media from ATCC, supplemented with Renal Epithelial Cell Growth Kit components (PCS-400-040) in a humidified atmosphere at 37 °C with 5% CO2.

Techniques: Expressing, Incubation, Derivative Assay