Hacat

Manufactured by Cosmo Bio

Sourced in Japan

HaCaT is a spontaneously immortalized human keratinocyte cell line derived from adult human skin. It is a widely used in vitro model for studying various aspects of human keratinocyte biology and skin-related research.

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Lab products found in correlation

Hsc 3, by Japanese Collection of Research Bioresources Cell Bank (2 mentions) Hsc 2, by Japanese Collection of Research Bioresources Cell Bank (2 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions) Hacat, by Merck Group (1 mentions) F12 medium, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)

4 protocols using hacat

Cell Line Authentication and Cultivation

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We obtained the human cervical dysplasia cell line W12 (RRID:CVCL_T290, clone 20,863), which contains HPV16 episomes, from Drs. Paul Lambert, Tomomi Nakahara, and Iwao Kukimoto.¹⁹ This cell line was authenticated by STR profiling and was cultured as previously reported.¹¹We obtained HaCaT (RRID:CVCL_0038), which is a spontaneously transformed immortal keratinocyte cell line derived from human adult male skin,¹⁶ from Cosmo Bio (Tokyo, Japan) and cultured it as previously reported.¹¹The human AR‐positive prostate cancer cell line LNCaP (RRID:CVCL_ 0395), which was authenticated by STR profiling, was gifted by Prof. Atsushi Mizokami, Department of Urology, Graduate School of Medical Sciences, Kanazawa University and was used for Western blot or RT‐PCR analysis as positive controls.
Mycoplasma infections were detected regularly and all experiments were performed under mycoplasma‐free conditions.

Matsumoto T., Suzuki T., Nakamura M., Yamamoto M., Iizuka T., Ono M., Kagami K., Kasama H., Kanda T., Sakai Y., Iwadare J., Matsuoka A., Kayahashi K., Wakae K., Muramatsu M., Kyo S., Yamamoto Y., Mizumoto Y., Daikoku T, & Fujiwara H. (2023). Androgen promotes squamous differentiation of atypical cells in cervical intraepithelial neoplasia via an ELF3‐dependent pathway. Cancer Medicine, 12(9), 10816-10828.

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Cell Culture Protocol for Cervical Dysplasia and Cancer

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The human cervical dysplasia cell line W12 (RRID:CVCL_T290, clone 20863),²² which was authenticated using short tandem repeat (STR) profiling, was gifted by Drs Paul Lambert, Tomomi Nakahara, and Iwao Kukimoto and used in this study. This cell line contains HPV16 episomes. W12 cells were cultured at 37°C under 5% CO₂ in F medium composed of three parts F‐12 medium (Sigma‐Aldrich) and one part DMEM (Sigma‐Aldrich) supplemented with 5% FBS (Sigma‐Aldrich), 0.4 μg/ml hydrocortisone, 5 μg/ml insulin, 8.4 ng/ml cholera toxin, 24 μg/ml adenine, 10 ng/ml epidermal growth factor (EGF), 100 g/ml streptomycin, and 100 IU/ml penicillin.
The immortalized human keratinocytes cell line HaCaT (RRID:CVCL_0038) was purchased from Cosmo Bio. HaCaT cells were cultured at 37°C under 5% CO₂ in DMEM (Sigma‐Aldrich) or calcium‐free DMEM (Sigma‐Aldrich) supplemented with 10% FBS (Sigma‐Aldrich), 100 g/ml streptomycin, and 100 IU/ml penicillin.
The human cervical squamous carcinoma cell line C33A (RRID:CVCL_1094), which was authenticated by STR profiling, was used in this study. C33A cells (HPV negative) were cultured at 37°C under 5% CO₂ in DMEM (Sigma‐Aldrich) supplemented with 10% FBS (Sigma‐Aldrich), 100 g/ml streptomycin, and 100 IU/ml penicillin.

Matsumoto T., Iizuka T., Nakamura M., Suzuki T., Yamamoto M., Ono M., Kagami K., Kasama H., Wakae K., Muramatsu M., Horike S., Kyo S., Yamamoto Y., Mizumoto Y., Daikoku T, & Fujiwara H. (2022). FOXP4 inhibits squamous differentiation of atypical cells in cervical intraepithelial neoplasia via an ELF3‐dependent pathway. Cancer Science, 113(10), 3376-3389.

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Cell line cultivation for cancer research

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The human oral squamous cell carcinoma cell lines, HSC-2 (JCRB0622: oral squamous carcinoma cells derived from weak gingivae) and HSC-3 (JCRB0623: oral squamous carcinoma cells derived from highly advanced tongue cancer) (JCRB Cell Bank, Tokyo, Japan), human cervical adenocarcinoma cells (HeLa) (TKG0331, Cell Resource Center for Biomedical Research in Tohoku University, Miyagi, Japan), and the normal human keratinized epithelial cell line, HaCaT (Cosmo Bio Co., Ltd., Tokyo, Japan), were used in the present study. All cell lines were authenticated using a short-tandem repeat analysis. All cells were cultured in low glucose Dulbecco’s modified Eagle’s medium (DMEM) (041-29775, Fujifilm-Wako Pure Medical Corporation, Miyazaki, Japan) supplemented with 10% heat-inactivated fetal bovine serum (Life Technologies, New York, NY, USA), 100 mg/mL streptomycin, and 100 U/mL penicillin (Fujifilm-Wako Pure Medical Corporation, Miyazaki, Japan) at 37 °C in a humidified atmosphere with 5% CO₂.

Liu S., Washio J., Sato S., Abiko Y., Shinohara Y., Kobayashi Y., Otani H., Sasaki S., Wang X, & Takahashi N. (2022). Rewired Cellular Metabolic Profiles in Response to Metformin under Different Oxygen and Nutrient Conditions. International Journal of Molecular Sciences, 23(2), 989.

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Oral Squamous Carcinoma Cell Lines

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Human squamous carcinoma cell-derived strains; i.e., HSC-2 (JCRB0622: oral squamous carcinoma cells derived from weak gingivae) and HSC-3 (JCRB0623: oral squamous carcinoma cells derived from highly advanced tongue cancer) (JCRB Cell Bank, Tokyo, Japan) cells, and a normal human keratinized epithelial cell line, HaCaT (Cosmo Bio Co., Ltd., Tokyo, Japan), were used. All cell lines were authenticated by means of short-tandem repeat (STR) analysis.

Shinohara Y., Washio J., Kobayashi Y., Abiko Y., Sasaki K, & Takahashi N. (2021). Hypoxically cultured cells of oral squamous cell carcinoma increased their glucose metabolic activity under normoxic conditions. PLoS ONE, 16(10), e0254966.

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