H 6016

Manufactured by Cell Biologics

The H-6016 is a laboratory equipment product. It is a microplate incubator that maintains temperature and environmental conditions for cell culture and other biological applications. The core function of the H-6016 is to provide a controlled environment to support the growth and development of cells or other biological samples.

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Lab products found in correlation

Mab218, by R&D Systems (1 mentions) Pd98059, by Cell Signaling Technology (1 mentions) M2267, by Cell Biologics (1 mentions) Mab003, by R&D Systems (1 mentions) B7651, by Merck Group (1 mentions) U0126, by Cell Signaling Technology (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Cc 2553, by Lonza (1 mentions) Human il 6, by Toray (1 mentions) Human il 17a, by Thermo Fisher Scientific (1 mentions) Human tnf α, by Thermo Fisher Scientific (1 mentions)

2 protocols using h 6016

Isolation and Culture of Cardiac Fibroblasts

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Human cardiac fibroblasts were prepared as follows: right atrial biopsies were weighed, minced into 1–2 mm³ pieces, and placed in 6-cm dishes. Human cardiac fibroblasts were grown and maintained in DMEM (11995- 065, Gibco) supplemented with 20% fetal bovine serum (FBS, 10500, Hyclone) and 1% penicillin–streptomycin (15140-122, Gibco), in a humidified atmosphere at 37 °C and 5% CO₂. The medium was renewed every 2–3 days. At 80–90% confluence, cells were passaged using standard trypsinization techniques. This protocol was also used to isolate fibroblasts from mouse atria, ventricles and kidneys. Human primary kidney fibroblasts (H-6016; CellBiologics) were cultured in specific medium (M2267, CellBiologics). All experiments were carried out at low cell passage (< P4) and cells were cultured in serum-free media for 16 h before treatment.
Antibodies and inhibitors used in this study were as follows: IgG type 2a (MAB003, R&D Systems), anti-IL-11 antibody (MAB218, R&D Systems), anti- IL11RA antibody (MAB1977, R&D Systems), brefeldin A (B7651, Sigma-Aldrich), cycloheximide (C1988, Sigma-Aldrich), PD98059 (9900, Cell Signaling), and U0126 (9930, Cell Signaling).

Schafer S., Viswanathan S., Widjaja A.A., Lim W.W., Moreno-Moral A., DeLaughter D.M., Ng B., Patone G., Chow K., Khin E., Tan J., Chothani S.P., Ye L., Rackham O.J., Ko N.S., Sahib N.E., Pua C.J., Zhen N.T., Xie C., Wang M., Maatz H., Lim S., Saar K., Blachut S., Petretto E., Schmidt S., Putoczki T., Guimarães-Camboa N., Wakimoto H., van Heesch S., Sigmundsson K., Lim S.L., Soon J.L., Chao V.T., Chua Y.L., Tan T.E., Evans S.M., Loh Y.J., Jamal M.H., Ong K.K., Chua K.C., Ong B.H., Chakaramakkil M.J., Seidman J.G., Seidman C.E., Hubner N., Sin K.Y, & Cook S.A. (2017). IL-11 is a crucial determinant of cardiovascular fibrosis. Nature, 552(7683), 110-115.

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Renal Cell Responses to Cytokines

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Human primary kidney cells used in the experiments include renal proximal tubule epithelial cells (RPTEC; CC-2553, Lonza, Switzerland), human renal glomerular microvascular endothelial cells (HRGEC; ACBRI128, Cell Systems, Kirkland, WA), kidney fibroblasts (HKF; H-6016, Cell Biologics, Chicago, IL), and human renal mesangial cells (HRMC; #4200, ScienCell, Carlsbad, CA). The cells were plated in 96-well plates (1 × 10 4 cells/well) and stimulated with some combination of human IL-6 (100 ng/ml; Toray Industries) plus human soluble IL-6Rα (100 ng/ml; PeproTech), human IL-17A (50 ng/ml; PeproTech), recombinant human ORM1 (1,000 ng/ml; Prospecbio, Rehovot, Israel), and human TNFα (50 ng/ml; PeproTech) for 3, 6 or 24 hours after serum starvation. Soluble IL-6Rα was added, because non-immune cells normally express only IL-6 signal-transducing receptor subunit gp130. Recombinant ORM1 was inactivated by incubating at 100°C for 10 min. The cells were harvested, and total RNA was prepared for real-time PCR. For mechanistic analysis, RPTEC were immortalized using SV40 large T antigen. Immortalized RPTEC showed a similar response to primary RPTEC following cytokine stimulation (Supplementary Fig. 1). Human hepatoma Hep3B cells were stimulated overnight with IL-6/IL-6Rα, IL-17 or their combination.

Higuchi H., Kamimura D., Jiang J.J., Atsumi T., Iwami D., Hotta K., Harada H., Takada Y., Kanno-Okada H., Hatanaka K.C., Tanaka Y., Shinohara N, & Murakami M. (2020). Orosomucoid 1 is involved in the development of chronic allograft rejection after kidney transplantation. International immunology, 32(5).

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