Defined keratinocyte sfm

Manufactured by Thermo Fisher Scientific

Sourced in United States

Defined Keratinocyte-SFM is a serum-free, defined medium formulated for the growth and propagation of normal human epidermal keratinocytes. The medium contains essential growth factors and supplements required for the culture of keratinocytes in a defined, serum-free environment.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Keratinocyte-SFM (234 citations) Defined Keratinocyte-SFM medium (10 citations) Defined keratinocyte-SFM (1×) (8 citations) Defined keratinocyte-SFM growth supplement (4 citations) Defined Keratinocyte SFM (DKSFM) (2 citations) Defined keratinocyte-SFM (1X) (2 citations) Defined keratinocyte serum-free medium (K-SFM), (2 citations)

59 protocols using defined keratinocyte sfm

Isolation and Cultivation of Prostate Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Prostate tissues of HP patients were collected and washed three times with PBS solution. After that, tissues were digested on a shaker in a Defined Keratinocyte-SFM (Thermo Fisher, Waltham, MA, USA) supplemented by Collagenase IV (Abbexa, Cambridge, UK) 200 U/mL for 30 min at 37 °C. The suspension was filtered through mesh to separate single cells. Cell pellets were collected using centrifugation at 200 g for 10 min, then the cells were resuspended and cultured in a Defined Keratinocyte-SFM (Thermo Fisher, Waltham, MA, USA) supplemented with bovine pituitary extract and 5 mM epidermal growth factor (EGF) at 37 °C with 5% CO₂.
LNCap cells were propagated using RPMI 1640 (Thermo Fisher, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS), L-glutamine (2 mM), penicillin (100 U/mL), and streptomycin (100 mg/mL), at 37 °C with 5% CO₂.

Wang G.C., Huang T.R., Wang K.Y., Wu Z.L., Xie J.B., Zhang H.L., Yin L., Tang W.L, & Peng B. (2021). Inflammation induced by lipopolysaccharide advanced androgen receptor expression and epithelial-mesenchymal transition progress in prostatitis and prostate cancer. Translational Andrology and Urology, 10(11), 4275-4287.

+ Open protocol

+ Expand

NSCLC and Normal Lung Cell Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols

NSCLC cell lines, including A549, H1975, H1703, mouse LLC (LL/2) cell, non-cancerous HEK293FT (293FT), and human umbilical vein endothelial (HUVEC) cells were obtained from the Cell Bank of Shanghai Institutes of Biological Sciences (Shanghai, China) or ATCC, and cultured in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO) medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (penicillin 100 U/ml and streptomycin 10 μg/ml) or LSGS-supplemented medium 200PRF (for HUVEC, GIBCO). Primary normal lung epithelial and primarily cultured stage III LUAD cell (LC1) were cultured in Defined Keratinocyte SFM (GIBCO) supplemented with L-glutamine, EGF (20 ng/ml), basic-FGF (10 ng/ml), 2% B27, penicillin/streptomycin, and amphotericin B (0.25 mg/ml)^{56 (link),57 (link)}. All cell lines were authenticated by short tandem repeat (STR) fingerprinting at Medicine Laboratory of Forensic Medicine Department of Sun Yat-Sen University (SYSU) (Guangzhou, China), and were tested to be free of mycoplasma contamination.

Liu L., Tao T., Liu S., Yang X., Chen X., Liang J., Hong R., Wang W., Yang Y., Li X., Zhang Y., Li Q., Liang S., Yu H., Wu Y., Guo X., Lai Y., Ding X., Guan H., Wu J., Zhu X., Yuan J., Li J., Su S., Li M., Cai X., Cai J, & Tian H. (2021). An RFC4/Notch1 signaling feedback loop promotes NSCLC metastasis and stemness. Nature Communications, 12, 2693.

+ Open protocol

+ Expand

Establishment and Characterization of Human Oral Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

HGK (PCS-200–014) and human primary gingival fibroblasts (CRL-2014) were purchased from the American Type Culture Collection and cultured in KGM-Gold™ BulletKit™ (Lonza Japan, Tokyo, Japan) and KBM Fibro Assist (KOHJIN BIO, Saitama, Japan), respectively. Human oral keratinocytes (HOK) isolated from the oral mucosa were purchased from ScienCell Research Laboratories (San Diego, CA) and cultured in KGM-Gold™ BulletKit™. A Simian virus-40 antigen immortalized human gingival keratinocyte cell line (OBA-9), which was generated by Prof. Shinya Murakami [16 (link)], was maintained in Defined Keratinocyte-SFM (Gibco, Carlsbad, CA, USA). Normal human lung fibroblasts were provided by the Japanese Collection of Research Bioresources Cell Bank. The HaCaT immortalized human skin keratinocyte cell line was obtained from N. E. Fusenig (German Cancer Research Center, Heidelberg, Germany) and cultured in DMEM supplemented with 10% FBS. Human primary epidermal keratinocytes (HEK) were purchased from Lifeline Cell Technology (Frederick, MD, USA) and maintained in HuMedia-KG2 (Kurabo Industries, Osaka, Japan). In some experiments, HGK were stimulated with various concentrations of 3p-hpRNA using Lipofectamine RNAiMAX transfection reagent, according to the manufacturer’s instructions.

Shikama Y., Kurosawa M., Furukawa M., Kudo Y., Ishimaru N, & Matsushita K. (2022). The Priming Potential of Interferon Lambda-1 for Antiviral Defense in the Oral Mucosa. Inflammation, 45(3), 1348-1361.

+ Open protocol

+ Expand

Cell Culture of Various Cancer Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human embryonic kidney (HEK) 293T cells, CNE2, and HeLa-Bx1 cells were maintained in Dulbecco's Minimal Essential Medium (Gibco) supplemented with 10% fetal bovine serum (FBS) and 1% P/S. The EBV-positive NPC cell line C666-1 and the Burkitt's lymphoma lines Mutu III, Mutu I and DG-75 were grown in RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS) and 1% P/S. The hTERT immortalized NP epithelial cell lines NP361-hTERT-EBV and NP460-hTERT-EBV were grown in a 1:1 mixture of Defined Keratinocyte-SFM (Gibco) and Epilife™ medium (Gibco) supplemented with 1% P/S. The EBV-positive and -negative gastric cancer cell line AGS-Bx1 and AGS, respectively, were cultured in F-12K Nutrient Mixture (Gibco) supplemented with 10% fetal bovine serum (FBS) and 1% P/S. Cells were cultured at 37°C with 5% CO₂.

Verhoeven R.J., Tong S., Mok B.W., Liu J., He S., Zong J., Chen Y., Tsao S.W., Lung M.L, & Chen H. (2019). Epstein-Barr Virus BART Long Non-coding RNAs Function as Epigenetic Modulators in Nasopharyngeal Carcinoma. Frontiers in Oncology, 9, 1120.

+ Open protocol

+ Expand

Cell Culture Protocols for NPC and Lung Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

The immortalized nasopharyngeal epithelial NP69 cell line was cultured in keratinocyte serum-free medium (SFM) (Invitrogen, Carlsbad, USA) with the SFM-Growth supplement. The immortalized nasopharyngeal epithelial NP460 cell line was grown in a 1:1 ratio of Defined Keratinocyte-SFM (Gibco, NY, USA) supplemented with growth factors. The human NPC cell lines, including HK1, C666–1, CNE1, CNE2, CNE1-LMP1, HNE1, HNE2, HONE1, 5–8F, 6-10B, SUNE1 and lung carcinoma cell lines, including A549 (ATCC No.CCL-185) and H1299 (ATCC No.CRL-5803) cell lines were grown in RPMI 1640 media supplemented with 10% v/v heat-inactivated fetal bovine serum (FBS), 1% w/v glutamine and 1% w/v antibiotics. All the cell lines involved were cultured at 37 °C in a humidified incubator containing 5% CO₂.

Luo X., Li N., Zhao X., Liao C., Ye R., Cheng C., Xu Z., Quan J., Liu J, & Cao Y. (2019). DHRS2 mediates cell growth inhibition induced by Trichothecin in nasopharyngeal carcinoma. Journal of Experimental & Clinical Cancer Research : CR, 38, 300.

+ Open protocol

+ Expand

Culturing Human Cervical Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human cervical cancer cells (HeLa-cells) were a kind gift provided from Professor Y.W. Lin, Department of Microbiology and Immunology, Taipei, Taiwan. The human normal endocervical cells (End1-cells/E6E7, CRL-2615^®) and human normal ectocervical cells (Ect1-cells/E6E7, CRL-2614^®) were bought commercially from American Type Culture Collection (ATCC^®, Manassas, VA, USA). HeLa cells were cultured in 90% Dulbecco’s Modified Eagle Medium (pH_o = 7.4) with high glucose (25 mM) containing 17.9 mM NaHCO₃, 10% FBS (fetal bovine serum), 1% non-essential amino acids, 1% sodium pyruvate, and 1% antibiotic antimycotic solution (100 units/mL penicillin, 10 mg/mL streptomycin sulfate, and 0.025 mg/mL amphotericin-B). End1 and Ect1 cells were cultured in Defined Keratinocyte-SFM (10744-019, Gibco^TM, San Jose, CA USA) containing 1% antibiotic antimycotic solution. All cells were cultured at 37 °C/5% CO₂, pH_o = 7.4, and the culture media was renewed every 2–3 days. When the cells reached 70–80% in growth density, they were routinely subcultured and further experiments were conducted.

Loh S.H., Tsai Y.T., Huang S.F., Yu T.C., Kuo P.C., Chao S.C., Chou M.F., Tsai C.S, & Lee S.P. (2020). Effects of Andrographolide on Intracellular pH Regulation, Cellular Migration, and Apoptosis in Human Cervical Cancer Cells (Running Tittle: Effects of Andrographolide on pH Regulators and Apoptosis in Cervical Cancer). Cancers, 12(2), 387.

+ Open protocol

+ Expand

Culturing Human Nasopharyngeal Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human NPC cell lines, HK1, HK1-EBV, HONE1, and HONE1-EBV, were generously provided by Professor Sai Wah Tsao from the University of Hong Kong. Cells were cultured in RPMI-1640 medium (Cat: 11875500, Gibco, Grand Island, USA) supplemented with 10% fetal bovine serum (FBS; Cat: 04-001-1, BI, Kibbutz Beit-Haemek, Israel). The immortalized human nasopharyngeal epithelial cell lines, NP460hTERT and NP460hTERT-EBV, were generously provided by Professor Sai Wah Tsao from the University of Hong Kong.Cells were cultured in a 1:1 mixture of Defined Keratinocyte-SFM and EpiLife medium (Cat: 10744019 and MEPI500CA, Gibco). Cells were cultured in 37 °C incubators with 5% CO₂.

Li Y., Shi F., Hu J., Xie L., Zhao L., Tang M., Luo X., Ye M., Zheng H., Zhou M., Liu N., Bode A.M., Fan J., Zhou J., Gao Q., Qiu S., Wu W., Zhang X., Liao W, & Cao Y. (2021). Stabilization of p18 by deubiquitylase CYLD is pivotal for cell cycle progression and viral replication. NPJ Precision Oncology, 5, 14.

+ Open protocol

+ Expand

Cell Line Characterization and Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Cal27 and FaDu cell lines were obtained from the ATCC (United States); the HSC-3 cell line was obtained from the JCRB Cell Bank (Japanese); the SCC4 cell line was purchased from the BeNa Culture Collection (BNCC, China); and the UM-SCC1 and human normal oral keratinocyte (NOK) cell lines were obtained from the State Key Laboratory of Oral Diseases. All cancer cell lines were authenticated by STR analysis. Cal27, FaDu, SCC4, UM-SCC1 and HSC-3 cells were cultured in DMEM (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum, while NOK cells were cultured in defined keratinocyte SFM (Gibco, Grand Island, NY, USA) with corresponding growth supplement. All cells were maintained at 37 °C in a humidified atmosphere with 5% CO₂. For cellular intervention involving inhibitors or activators, all reagents were applied at specific final concentrations, which were diluted with dimethyl sulfoxide (DMSO). An equivalent volume of DMSO was added as a negative control. The final concentrations are indicated as described in the figure legends.

Liu Y., Sun Y., Yang J., Wu D., Yu S., Liu J., Hu T., Luo J, & Zhou H. (2024). DNMT1-targeting remodeling global DNA hypomethylation for enhanced tumor suppression and circumvented toxicity in oral squamous cell carcinoma. Molecular Cancer, 23, 104.

+ Open protocol

+ Expand

Establishment of Rat SCC Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

The SD rat SCC cell lines Rca-T (21 (link)) and Rca-B (22 (link)) were cultured in DMEM (Gibco, USA) supplemented with 10% FBS, 100 units/mL penicillin and 100 mg/mL streptomycin at 37°C in a humidified atmosphere of 5% CO₂. The primary culture came from buccal mucosa of male Sprague-Dawley (SD) rats, aged 4-5 weeks. The tissue was cut to 0.2 cm×0.2 cm pieces, then they were incubated in 5 mL 0.8% dispase II (Roche, Switzerland) solution at 4°C overnight. Epithelium was isolated from basal lamina, and put into 3 mL 0.25% trypsin for 5 minutes. The suspension was centrifuged in low speed and supernatant was decanted. The deposition was resuspended and incubated with Defined Keratinocyte SFM (Gibco, USA) in a 6 cm dish.

Zhang T., Zhu X., Sun Q., Qin X., Zhang Z., Feng Y., Yan M, & Chen W. (2021). Identification and Confirmation of the miR-30 Family as a Potential Central Player in Tobacco-Related Head and Neck Squamous Cell Carcinoma. Frontiers in Oncology, 11, 616372.

+ Open protocol

+ Expand

Colony Formation Assay for Stem and Progenitor Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells sorted by flow cytometry or bulk population single cells were plated in methylcellulose (Methocult from Stem Cell Technology, France) mixed with one of the following media: Stem cell medium I: 20 ng/ml EGF (PeproTech, USA), 10 ng/ml bFGF (PeproTech, USA), 1× B27 (Gibco) and 1× PS (Lonza) in 1× Defined Keratinocyte-SFM (Gibco); Stem Cell medium II: 20 ng/ml EGF, 20 ng/ml bFGF, 1× B27, 1× PS and 1× Glutamax (Gibco) in DMEM-F12 (Gibco); Pancreatic medium: Pancreatic culture media with pancreatic cell culture supplement and PS and RPMI medium: 10% FCS, 1× PS and 1× Glutamax in RPMI 1640 (Lonza) according to manufacturers' protocol. In brief, single cell suspensions were added to media with Methocult, mixed and seeded at a density of 1000 cells/well in a non-adhesive 24 well plate (Grenier Bio-One, Germany). Two weeks post plating, the colonies were stained with 150 µl 0.4 mg/ml Thiazolyl Blue Tetrazolium Bromide (MTT) (Sigma-Aldrich) at 37°C for 4 hours or overnight. Uniform colonies of minimum 50 µm were scanned using the GelCount machine and the number and sizes were quantified using GelCount software (both from Oxford Optronix, UK).

Wennerström A.B., Lothe I.M., Sandhu V., Kure E.H., Myklebost O, & Munthe E. (2014). Generation and Characterisation of Novel Pancreatic Adenocarcinoma Xenograft Models and Corresponding Primary Cell Lines. PLoS ONE, 9(8), e103873.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!