Cc 3151

Manufactured by Lonza

Sourced in Switzerland

The CC-3151 is a reliable lab equipment product designed for general laboratory use. It serves as a core function in various research and testing applications. The product specifications and technical details are available upon request.

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Lab products found in correlation

Cc 4136, by Lonza (5 mentions) Lenti x 293t cells, by Takara Bio (3 mentions) Cc 4009, by Lonza (3 mentions) Mcf 10a, by Lonza (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Mcf 7, by Thermo Fisher Scientific (2 mentions) Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (2 mentions) Mir 590 3p and mir 200a 3p mimics, by Merck Group (2 mentions) Cc 4031g, by Lonza (2 mentions) Cc 4021g, by Lonza (2 mentions) Mcf 7, by American Type Culture Collection (2 mentions) Mcf 10a, by American Type Culture Collection (2 mentions) Skbr3, by American Type Culture Collection (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Sk br 3, by Korean Cell Line Bank (1 mentions) Sk br 3, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Cc 4176, by Lonza (1 mentions) Mcf 7, by Korean Cell Line Bank (1 mentions)

16 protocols using cc 3151

Human Breast Cancer Cell Lines Protocol

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Human breast cancer (MCF7, MDA-MB-231, T47D, and SK-BR-3) cells were purchased from the Korean Cell Line Bank (Seoul, Korea). Normal human breast (MCF10A) cells and primary human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). MCF7, T47D, and SK-BR-3 cells were grown in the RPMI-1640 medium (Gibco, Grand Island, NY, USA), and MDA-MB-231 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) containing 10% fetal bovine serum (FBS, Sigma, Darmstadt, Germany) with 100 mg/mL penicillin and streptomycin (P/S, GenDEPOT, Barker, TX, USA). MCF10A cells were grown in the mammary epithelial basal medium (CC-3151; Lonza, Basel, Switzerland) containing supplements (CC-4136; Lonza). HUVECs were grown in endothelial basal medium-2 (CC-3156; Lonza) containing supplements (CC-4176; Lonza) according to the manufacturer’s instructions.

Jo M.J., Kim B.G., Kim W.Y., Lee D.H., Yun H.K., Jeong S., Park S.H., Kim B.R., Kim J.L., Kim D.Y., Lee S.I, & Oh S.C. (2021). Cannabidiol Suppresses Angiogenesis and Stemness of Breast Cancer Cells by Downregulation of Hypoxia-Inducible Factors-1α. Cancers, 13(22), 5667.

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Scaffold-Aided Keratinocyte Differentiation

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In the current experiment, human cutaneous basal cells were used to investigate whether scaffolds have the potential to promote cell differentiation toward mature keratinocytes. For this proposes, human basal cells were obtained from the Stem Cell Research Center Cell Baking (Tabriz University of Medical Sciences) that obtained previously from human skin samples [4 (link)]. To expand the BCCs, we used epithelial cell growth basal medium (Cat no: CC-3151; Lonza) containing growth factor cocktail such as keratinocyte growth factor, insulin-like growth factor, heparin, hydrocortisone, fibroblast growth factor, epidermal growth factor, ascorbic factor (CC-4136; Lonza), 1% Pen-Strep (Gibco) and 10% fetal bovine serum (Gibco). Cells were detached at 70–80% confluence by using an enzymatic solution Trypsin-EDTA (0.25%; Gibco). Cells at passages 3–5 were used in the various analyses. The exhausted medium was replenished every 3–4 days.

Mohammadzadeh L., Rahbarghazi R., Salehi R, & Mahkam M. (2019). A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering. Journal of Biological Engineering, 13, 79.

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Lentivirus Production and Cell Culture

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LentiX-293T cells (Clontech) were cultured in DMEM (with high glucose, glutamine and sodium pyruvate) with 5% FBS and 1× non-essential amino acid. LentiX-293T cells were used to make lentivirus. Ba/F3 cells are murine pro-B suspension cells that depend on exogenous IL-3 for cell survival. Growth medium for Ba/F3 cells was advanced RPMI with 1× GlutaMAX, 5% FBS and 1 ng/ml mouse IL-3. Assay medium for Ba/F3 was growth medium without IL-3. MCF10A cells are human non-tumorigenic mammary epithelial cells that depend on exogenous EGF and insulin for proliferation. Growth medium for MCF10A cells was DMEM/F12 medium with 5% HS, 20 ng/ml EGF, 0.5 mg/ml hydrocortisone, 100 ng/ml cholera toxin, 10 μg/ml insulin, 1× Pen/strep. Assay medium for MCF10A cells was MEBM basal medium (Lonza #CC-3151) with 100 ng/ml cholera toxin and 52 ng/ml bovine pituitary extract (BPE) (Lonza #CC-4009).

Ng P.K., Li J., Jeong K.J., Shao S., Chen H., Tsang Y.H., Sengupta S., Wang Z., Bhavana V.H., Tran R., Soewito S., Minussi D.C., Moreno D., Kong K., Dogruluk T., Lu H., Gao J., Tokheim C., Zhou D.C., Johnson A.M., Zeng J., Ip C.K., Ju Z., Wester M., Yu S., Li Y., Vellano C.P., Schultz N., Karchin R., Ding L., Lu Y., Cheung L.W., Chen K., Shaw K.R., Meric-Bernstam F., Scott K.L., Yi S., Sahni N., Liang H, & Mills G.B. (2018). Systematic Functional Annotation of Somatic Mutations in Cancer. Cancer cell, 33(3), 450-462.e10.

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Immortalization of human cells

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Human primary BJ fibroblasts were cultured in DMEM with 10% FBS. Human mammary epithelial cells were cultured in MEGM mammary epithelial medium (Lonza CC-3151). BJ and HMEC cells were first retrovirally transduced to express ecotropic receptor and then modified with retrovirally transduced p53 shRNA, p16^INK4A shRNA, small T antigen, inducible ER-Ras^V12, and hTert as described (Elenbaas et al, 2001 (link); Voorhoeve & Agami, 2003 (link)). Retrovirus was made by calcium phosphate transfection of 293T cells and harvested after 48 h. BJ or HMEC cells were transduced with virus for 24 h and selected in medium containing 2–4 μg/ml blasticidin for 2–4 days before plating in soft agar.

Hong X., Nguyen H.T., Chen Q., Zhang R., Hagman Z., Voorhoeve P.M, & Cohen S.M. (2014). Opposing activities of the Ras and Hippo pathways converge on regulation of YAP protein turnover. The EMBO Journal, 33(21), 2447-2457.

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Culturing Human Breast Cancer and Kidney Cells

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Human breast cancer cell lines BT-474, MDA-MB-453, MDA-MB-231, and human embryonic kidney cells (HEK 293T) were grown and maintained in Dulbecco modified Eagle medium (DMEM) (Sigma, D5648) supplemented with 10% (v/v) fetal bovine serum (FBS) (Gibco, 10270) and 1% penicillin–streptomycin (Hyclone, 113-98-43810-74-0). Non-tumorigenic mammary epithelial (MCF-10A) cells were grown and maintained in complete mammary epithelial basal medium (MEBM) (Lonza, CC-3151). All cell lines were maintained at 37 °C, 5% CO₂, and 95% humidity in a CO₂ incubator.

Pani T., Rajput K., Kar A., Sharma H., Basak R., Medatwal N., Saha S., Dev G., Kumar S., Gupta S., Mukhopadhyay A., Malakar D., Maiti T.K., Arimbasseri A.G., Deo S.V., Sharma R.D., Bajaj A, & Dasgupta U. (2021). Alternative splicing of ceramide synthase 2 alters levels of specific ceramides and modulates cancer cell proliferation and migration in Luminal B breast cancer subtype. Cell Death & Disease, 12(2), 171.

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Establishing Stable HT1080-FUCCI Cell Line

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HT1080 cells were obtained from ATCC and maintained in Minimum Essential Medium (MEM) alpha supplemented with 10% fetal bovine serum, 100 mg/ml streptomycin sulfate and 100 U/ml penicillin. To establish the stable expression of cell cycle markers (HT1080-FUCCI), HT1080-EYFP-53BP1 cells (19 (link)) were transduced with lentivirus carrying G1 [mCherry-hCdt1(30/120)] and S/G2 [AmCyan-hGeminin(1/110)] phase markers. Stable HT1080-FUCCI cells were selected using zeocin (1 μg/ml). To down-regulate the expression of RAD51, HT1080-FUCCI cells were transfected with a mammalian expression plasmid containing tetracycline-inducible Rad51 shRNA (20 (link)). Stable lines were selected using hygromycin (75 μg/ml). To down-regulate RAD51 protein levels, cells were cultured in the presence of doxycycline (DOX, 1 μg/ml) for 72 h. MCF10A cells stably expressing scrambled and Rad51 shRNA were kind gift from Dr Shiaw-Yih Lin (MD Anderson Cancer center) and were maintained in mammary epithelial basal medium (LONZA, CC-3151) as described previously (21 (link)). 4T1 cells were maintained as described previously (22 (link)). To down-regulate the expression of RAD51, 4T1 cells were transfected with a mammalian expression plasmid containing tetracycline-inducible Rad51 shRNA and selected using puromycin (1 μg/ml). All cells were maintained at 37°C in a humidified 5% CO₂ incubator.

Bhattacharya S., Srinivasan K., Abdisalaam S., Su F., Raj P., Dozmorov I., Mishra R., Wakeland E.K., Ghose S., Mukherjee S, & Asaithamby A. (2017). RAD51 interconnects between DNA replication, DNA repair and immunity. Nucleic Acids Research, 45(8), 4590-4605.

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Cell Culture and miRNA Transfection Protocol

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LentiX-293T cells (Clontech) were cultured in DMEM (with high glucose, glutamine and sodium pyruvate) with 5% fetal bovine serum and 1 × non-essential amino acid. MCF10A cells (ATCC® CRL-10317) were cultured in specific growth medium (DMEM/F12 medium with 5% HS, 20 ng/ml epidermal growth factor, 0.5 mg/ml hydrocortisone, 100 ng/ml cholera toxin, 10 μg/ml insulin, 1 × Pen/strep). Assay medium for MCF10A cells was MEBM basal medium (Lonza #CC-3151) with 100 ng/ml cholera toxin and 52 ng/ml bovine pituitary extract (Lonza #CC-4009).
HCT116 and KM12 colon carcinoma cell lines were purchased from MD Anderson Characterized Cell Line Core facility. Cell lines were confirmed using Short Tandem Repeat (STR) analysis and were tested and negative for mycoplasma contamination. Cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum. MiR-590-3p and miR-200a-3p mimics, and MISSION miRNA Negative Control 1 (HMC0002) were purchased from Sigma Aldrich. Cells were transfected with 50nM of miRNA mimics or miRNA Negative Control using Lipofectamine RNAiMAX reagent (Thermo Fisher).

Wang Y., Xu X., Maglic D., Dill M.T., Mojumdar K., Ng P.K., Jeong K.J., Tsang Y.H., Moreno D., Bhavana V.H., Peng X., Ge Z., Chen H., Li J., Chen Z., Zhang H., Han L., Du D., Creighton C.J., Mills G.B., Camargo F, & Liang H. (2018). Comprehensive Molecular Characterization of the Hippo Signaling Pathway in Cancer. Cell reports, 25(5), 1304-1317.e5.

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Single-Cell Barcoding for Transcriptomics

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Single-cell suspensions were pelleted at 400 x g for 5 min and washed once with 10 mL mammary epithelial basal medium (MEBM; Lonza CC-3151). For each sample, one million cells were aliquoted, washed a second time with 200 μL MEBM, and resuspended in 90 μL of a 200 nM solution containing equimolar amounts of anchor lipid-modified oligonucleotides (LMOs) and sample barcode oligonucleotides in phosphate buffered saline (PBS). Following a 5-minute incubation on ice with anchor-LMO/barcode, 10 uL of 2 μM co-anchor LMO in PBS was added to each sample (for a final concentration of 200 nM), and wells were mixed by gentle pipetting and incubated for an additional 5 min on ice. Following incubation, cells were washed twice in 200 μL PBS with 1% BSA and pooled together into a single 15 mL conical tube containing 10 mL PBS/1% BSA. All subsequent steps were performed on ice.

Murrow L.M., Weber R.J., Caruso J.A., McGinnis C.S., Phong K., Gascard P., Rabadam G., Borowsky A.D., Desai T.A., Thomson M., Tlsty T, & Gartner Z.J. (2022). Mapping hormone-regulated cell-cell interaction networks in the human breast at single-cell resolution. Cell systems, 13(8), 644-664.e8.

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Breast Cancer Cell Line Cultivation

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All the breast cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). The following cell lines were used: MCF‐10A cells, MCF‐7 cells, MDA‐MB‐468 cells, SK‐BR‐3 cells, MDA‐MB‐231 cells. MCF‐10A cells were cultured in Mammary Epithelial Cell Basal Medium (Lonza; CC3151) supplemented with 0.4% bovine pituitary extract (Lonza; CC4009G), 0.1% human epidermal growth factor (Lonza; CC4017G), 0.1% hydrocortisone (Lonza; CC4031G), 0.1% GA‐1000 (Lonza; CC4081G), and 0.1% insulin (Lonza; CC4021G). The other four cells were cultured in DMEM (Gibco‐BRL) with 10% fetal bovine serum (GIBCO‐BRL) and 1% penicillin/streptomycin (GIBCO‐BRL). All the cells were incubated in a humidified atmosphere of 5% CO₂/95% air at 37 °C. Subcultivation of the cell lines was performed using 0.25% trypsin and 5 × 10⁻³m methylenediaminetetraacetic acid (EDTA) (Gibco‐BRL Co., MD, USA).

Ye S., Li W., Wang H., Zhu L., Wang C, & Yang Y. (2021). Quantitative Nanomechanical Analysis of Small Extracellular Vesicles for Tumor Malignancy Indication. Advanced Science, 8(18), 2100825.

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Cell Line Characterization and Culture Conditions

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MCF7, MCF10A, MDA-MB-231, MDA-MB-436, MDA-MB-453 and SKBR3 were purchased from American Type Culture Collection (ATCC). HMEL was provided by Dr Robert Weinberg, Whitehead Institute. All cell lines were tested and free of mycoplasma contamination (MycoAlert Mycoplasma Detection Kit, Lonza). MCF7, MDA-MB-231, MDA-MB-436, MDA-MB-453 and SKBR3 cells were grown in DMEM supplemented with 10% foetal bovine serum (FBS), 100 units per ml penicillin and 100 μg ml⁻¹ streptomycin. MCF10A cells were cultured in defined mammary epithelial basal media (CC-3151, MEBM Bullet Kit, Lonza, Inc.) supplemented with 50 μl of 1 mg ml⁻¹ cholera toxin and 2.5% FBS. HMEL cells were cultured in mammary epithelial growth media (CC-3051, MEGM, Lonza, Inc.).

Gumireddy K., Li A., Kossenkov A.V., Sakurai M., Yan J., Li Y., Xu H., Wang J., Zhang P.J., Zhang L., Showe L.C., Nishikura K, & Huang Q. (2016). The mRNA-edited form of GABRA3 suppresses GABRA3-mediated Akt activation and breast cancer metastasis. Nature Communications, 7, 10715.

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