M2267

Manufactured by Cell Biologics

The M2267 is a high-performance cell culture incubator designed for maintaining optimal cell growth conditions. It features precise temperature, CO2, and humidity control to support a wide range of cell types. The incubator provides a stable and uniform environment to ensure consistent and reliable cell culture results.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Mab218, by R&D Systems (1 mentions) Pd98059, by Cell Signaling Technology (1 mentions) H 6016, 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) Mycoalert, by Lonza (1 mentions) Geneprint 10 system, by Promega (1 mentions) Fetal bovine serum (fbs), by Cell Biologics (1 mentions) Cc 2512, by Lonza (1 mentions)

5 protocols using m2267

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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Characterization of Diverse Cancer Cell Lines

Cited 4 times

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Antibodies and dilutions used are detailed in Supplementary Table 1. Human rPDGF-C was from Sigma (SRP3139). TSAE1 (TS/A-E1)^{47 (link)}, HRM1 (ref. ^{48 (link)}), EMT6 (ref. ^{49 (link)}) and F3II^{50 (link)} cells were obtained from L. Wakefield^{12 (link)} (National Cancer Institute, USA) with permission of C. De Giovanni, J. Zhao, S. Rockwell and D. Alonso, respectively. D2A1 and D2.OR cells were from A. Chambers’ laboratory stocks^{51 (link)} (London Health Sciences Centre, Canada). D2A1-m1 and D2A1-m2 metastatic sublines were generated in house^{13 (link)}. ZR-75-1 (CRL-1500), 10T1/2 (CCL-226), 3T3 (CRL-1658), 4T1 (CRL-2539), IMR90 (CCL-186), MRC5 (CCL-171) and HEK293T (CRL-3216) cells were from ATCC. AT-3 cells were provided by C. Paget and D. Soulard (Institute Pasteur de Lille). GFP-positive CAFs were from 4T1-TB BALB/c Ub-GFP mice^{52 (link)}. Young (BALB-5013 and C57-6013) and aged (A57-6013, two independent batches from 58–78-week-old mice) mouse primary lung fibroblasts (Cell Biologics) were cultured in fibroblast medium (M2267, Cell Biologics) until immortalized (see below), after which they were cultured in DMEM (10% FBS). All other cells were maintained in DMEM (10% FBS) unless otherwise stated. All cells were routinely checked for mycoplasma contamination (MycoAlert, Lonza). The identity of ZR-75-1 cells was confirmed by short tandem repeat (GenePrint 10 System, Promega).

Turrell F.K., Orha R., Guppy N.J., Gillespie A., Guelbert M., Starling C., Haider S, & Isacke C.M. (2023). Age-associated microenvironmental changes highlight the role of PDGF-C in ER+ breast cancer metastatic relapse. Nature Cancer, 4(4), 468-484.

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Culturing Primary Intestinal Fibroblasts

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For coculture demonstrations presented in Extended Data Fig. 5, we used primary mouse intestinal fibroblasts. For initial expansion from cryostorage, mouse intestinal fibroblasts were cultured in 75-cm² flasks according to the manufacturer’s protocols using complete fibroblast medium (M2267, Cell Biologics) supplemented with growth factors. For modeling intestinal fibrosis (Fig. 5), we used primary human intestinal fibroblasts. In all experiments, we used cells between passages 3 and 6.

Sutton A.J., Duval S.J., Tweedie R.L., Abrams K.R, & Jones D.R. (2000). Empirical assessment of effect of publication bias on meta-analyses. BMJ : British Medical Journal, 320(7249), 1574-1577.

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Cardiac Fibroblast and NIH3T3 Culture

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Cardiac fibroblasts (CFs; BALB/c mouse primary cardiac fibroblasts; Cell Biologics; BALB-5049) were cultured in fibroblast medium provided by Cell Biologics (M2267), which contained 0.5 mL fibroblasts growth factor (0.1% v/v), 0.5 mL hydrocortisone (0.1% v/v), 5 mL 100x antibiotic-antimycotic solution, 5 mL of 200 mM L-Glutamine, and 10% (v/v) fetal bovine serum. NIH3T3 cells (mouse embryonic fibroblasts; ATCC; CRL-1658) were cultured in high glucose Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher Scientific; 11965092) supplemented with 10% (v/v) fetal bovine serum (Atlanta Biologicals; S22660), 5 mL of 10 units/L penicillin, and 5 mL 10 mg/mL streptomycin (Thermo Fisher Scientific; 15140122). Prior to seeding CFs, all culture vessels were pre-coated with a gelatin-based coating solution (Cell Biologics; 6950) for one hour. Both cell lines were maintained under standard conditions of 5% CO₂ at 37°C and 95% humidity.

Patricelli C., Lehmann P., Oxford J.T, & Pu X. (2023). Doxorubicin-Induced Modulation of TGF-β Signaling Cascade in Mouse Fibroblasts: Insights into Cardiotoxicity Mechanisms. Research Square.

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Activating Human Breast Fibroblasts

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hMFs and hLFs were purchased from Cell Biologics (H-6071) and Lonza (CC-2512), respectively, and cultured in complete fibroblast medium (Cell Biologics; M2267). The primary cells were activated by culturing in TGF-β (5 ng/ml) for 48 h. Human breast fibroblasts isolated from breast cancer patients (hbCAFs) were purchased from Neuromics (CAF06) and cultured in MSC-GRO low serum, complete medium (SC00B1).

Sharma M., Turaga R.C., Yuan Y., Satyanarayana G., Mishra F., Bian Z., Liu W., Sun L., Yang J, & Liu Z.R. (2021). Simultaneously targeting cancer-associated fibroblasts and angiogenic vessel as a treatment for TNBC. The Journal of Experimental Medicine, 218(4), e20200712.

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