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The CRL1213 is a laboratory equipment product used for cell culture applications. It is designed to provide a controlled and regulated environment for the growth and maintenance of cells. The CRL1213 offers temperature, humidity, and gas (CO2) control to support optimal cell culture conditions.

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Lab products found in correlation

Cyquant cell proliferation assay, by Thermo Fisher Scientific (2 mentions) Opti mem, by Thermo Fisher Scientific (2 mentions) Crl 1658, by American Type Culture Collection (1 mentions) Dtx 880 multimode detector, by Beckman Coulter (1 mentions) Synergy plate reader, by Agilent Technologies (1 mentions) Raw264, by American Type Culture Collection (1 mentions) Tib 71, by American Type Culture Collection (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Hemin, by Thermo Fisher Scientific (1 mentions)

7 protocols using crl1213

1

Cell Culture Conditions for Fibroblasts

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Rat skin fibroblast FR (ATCC® CRL1213™) used in these experiments were cultured in complete growth medium containing Eagle’s Minimum Essential Medium (ATCC®), and 10 % fetal bovine serum (FBS) at 37°C in an atmosphere of 5% CO2 and 95% air. This cell line was used in all experiments except the microsomal assay. Mouse embryonic fibroblast cells (NIH/3T3 ATCC® CRL-1658™) used in these experiments were cultured in the complete growth medium containing Eagle’s Minimum Essential Medium (ATCC®), and 10% fetal bovine serum (FBS) at 37°C in an atmosphere of 5% CO2 and 95% air. This cell line was used in the microsomal assay.
Crosby H.A., Ihnat M, & Miller K.E. (2015). Evaluating the Toxicity of the Analgesic Glutaminase Inhibitor 6-Diazo-5-Oxo-L-Norleucine in vitro and on Rat Dermal Skin Fibroblasts. MOJ toxicology, 1(1), 00005.
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2

Assessing Fibroblast Cell Cycle by Flow Cytometry

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Rat fibroblasts FR (ATCC® CRL1213™) were plated at 30% confluency in 6 well plates and allowed to attach overnight. DON was added at the IC50 concentration of 232.5 μM as determined by the CyQUANT® Cell Proliferation Assay (Molecular Probes) for 2 hours in serum free media and then 10% Cosmic Calf™ Serum (Thermo Hyclone) was added. Cells were incubated at 37°C with 5% CO2. After 48 hours incubation, cells were fixed in 50% ethyl alcohol solution and stored at 4°C for up to a week. After fixation, pelleted cells were suspended and labeled with propidium iodide (25μg/mL) in phosphate buffered saline (PBS), pH 7.4, containing 0.1% Triton X-100, 1mg/ml sodium citrate, and 100μg/mL RNase A. Cells were incubated for 30 minutes at 37°C. Cells were subjected to flow cytometry using a side scatter plot versus fluorescence (535nm excitation/617nm emission).
Crosby H.A., Ihnat M, & Miller K.E. (2015). Evaluating the Toxicity of the Analgesic Glutaminase Inhibitor 6-Diazo-5-Oxo-L-Norleucine in vitro and on Rat Dermal Skin Fibroblasts. MOJ toxicology, 1(1), 00005.
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3

Evaluation of Anti-Cancer Agents in Fibroblasts

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Clear 96-well plates were seeded with FR (ATCC® CRL1213™) rat fibroblasts at 2 × 104 cells/ml. On day 4, complete growth media was aspirated, and 100 μl of the following treatments were added: 0, 2, 20, or 200 μM concentrations of 6-diazo-5-oxo-l-norleucine and 6 μMcamptothecin in OptiMem (Invitrogen) 3% FBS. After 48 hours incubation, the treatments were aspirated and 10 μL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was added to each well. The plates were incubated for 2 hours at 37°C, 95% air and 5% CO2, until purple dye was visible. Detergent (sodium dodecyl sulfate) 100 μL was added to each well and the plates were covered in aluminum foil and incubated in a dark area for 2 hours at room temperature. Absorbance and fluorescence were read at 485nm excitation/535nm emission. Results were obtained using a Multimode Detector DTX 880 (Beckman Coulter).
Crosby H.A., Ihnat M, & Miller K.E. (2015). Evaluating the Toxicity of the Analgesic Glutaminase Inhibitor 6-Diazo-5-Oxo-L-Norleucine in vitro and on Rat Dermal Skin Fibroblasts. MOJ toxicology, 1(1), 00005.
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4

Cell Proliferation Assay of Rat Fibroblasts

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Cell proliferation was analyzed using CyQUANT® Cell Proliferation Assay (Molecular Probes). Rat fibroblasts FR (ATCC® CRL1213™) were plated at a density of 3,000 cells per well into white 96-well plates with clear bottoms and grown for 24 hours to allow for complete attachment. The cells were exposed to 100 μl of one of the following concentrations of 6-diazo-5-oxo-L-norleucine- in OptiMem (Invitrogen): 0.10, 0.45, 1.30, 4.10, 12.30, 37, 111, 333 or 1000 μM for 4 hours. Serum (10% Cosmic calf, Hyclone, Logan UT) was added to the wells and the cells incubated for 44 hours at 37°C. Exposure media was removed and the cells were lysed by freezing at −80°C for 72 hours. The cells were thawed and exposed to a 1:400 dilution of CyQUANT®/lysis buffer for 60 minutes. Plate fluorescence (480nm excitation, 520nm emission) was read on a Synergy plate reader (BioTek).
Crosby H.A., Ihnat M, & Miller K.E. (2015). Evaluating the Toxicity of the Analgesic Glutaminase Inhibitor 6-Diazo-5-Oxo-L-Norleucine in vitro and on Rat Dermal Skin Fibroblasts. MOJ toxicology, 1(1), 00005.
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5

Rat Dermal Fibroblasts and Macrophages

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Rat dermal fibroblast cell line (FR, CRL-1213, ATCC, Manassas, VA) and macrophages (RAW 264.7, TIB-71, ATCC, Manassas, VA) were used for the in vitro studies.
Ma H., Lam P.K., Siu W.S., Tong C.S., Lo K.K., Koon C.M., Wu X.X., Li X., Cheng W., Shum W.T, & Leung P.C. (2021). Adipose Tissue-Derived Mesenchymal Stem Cells (ADMSCs) and ADMSC-Derived Secretome Expedited Wound Healing in a Rodent Model – A Preliminary Study. Clinical, Cosmetic and Investigational Dermatology, 14, 753-764.
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6

Rat Dermal Fibroblasts: Glucose and AGEs Effects

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Rat dermal fibroblasts (cat. no. CRL-1213; American Type Culture Collection) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) without any supplementary antibiotics in an incubator (37˚C; 5% CO2). Fibroblasts were cultured in DMEM containing 10% FBS under twelve different conditions (according to the groupings) for 72 h after 24 h of serum deprivation (0.5% FBS). All manipulations were performed in dim lighting, and the plates were wrapped in aluminum foil to protect them from the light. Cells were sub-cultured after they had grown to confluence. All experiments were performed in triplicate.
The cells were divided into the following groups: i) normal glucose (NG) (NG 1.0 g/l; Gibco; Thermo Fisher Scientific, Inc.); ii) NG + Hemin (Hemin 5 µM; MilliporeSigma); iii) NG + chromium mesoporphyrin (CrMP) (CrMP 20 µM; MilliporeSigma); iv) NG + Hemin + CrMP; v) AGEs + NG (AGEs 100 µg/ml; Gibco; Thermo Fisher Scientific, Inc.); vi) AGEs + NG + Hemin; vii) AGEs + NG + CrMP; viii) AGEs + NG + Hemin + CrMP; ix) AGEs + HG (HG 4.5 g/l); x) AGEs + HG + Hemin; xi) AGEs + HG + CrMP; and xii) AGEs + HG + Hemin + CrMP group (9-11 (link)).
Li Q., Liang S., Lai Q., Shen L., Zhang Y, & Guo R. (2021). Heme oxygenase-1 alleviates advanced glycation end product-induced oxidative stress, inflammatory response and biological behavioral disorders in rat dermal fibroblasts. Experimental and Therapeutic Medicine, 22(5), 1212.
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7

Rat Bone Marrow MSC Isolation

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BM-MSCs were obtained from the femurs of adult Sprague-Dawley rats and cultured in DMEM-F12 supplemented with 10% heat inactivated FBS, 1% Glutamine, and 1% Penicillin/Streptomycin. MSCs were characterized by flow cytometry at passage 2 and used for a maximum of 5 passages. Cell lines, including nontransformed rat intestinal epithelial IEC-6 cells (CRL-1592, passage 13) and primary rat fibroblast (FB) cells (CRL-1213), were obtained from the American Type Culture Collection. IEC-6 cells used in all experiments were at or before the 20th passage.
Chen H., Min X.H., Wang Q.Y., Leung F.W., Shi L., Zhou Y., Yu T., Wang C.M., An G., Sha W.H, & Chen Q.K. (2015). Pre-activation of mesenchymal stem cells with TNF-α, IL-1β and nitric oxide enhances its paracrine effects on radiation-induced intestinal injury. Scientific Reports, 5, 8718.
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