Epithelial cell medium

Manufactured by ScienCell

Sourced in United States

Epithelial Cell Medium is a specialized culture medium formulated to support the growth and maintenance of epithelial cells in vitro. It provides the necessary nutrients and growth factors required for the optimal proliferation and differentiation of epithelial cell lines.

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27 protocols using epithelial cell medium

Gallbladder Cancer Cell Culture Protocol

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GBC-SD and SGC-996 cells were obtained from The Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Wuhan, China) and were cultured in RPMI-1640 medium supplemented with 10% FBS, 100 µg/ml streptomycin and 100 U/ml penicillin (all from Gibco; Thermo Fisher Scientific, Inc.). Normal human intrahepatic biliary epithelial cells (HIBECs) were obtained from ScienCell Research Laboratories, Inc. and were incubated using epithelial cell medium (ScienCell Research Laboratories, Inc.). Cells were incubated at 37°C with 5% CO₂. TRAIL was obtained from PeproTech, Inc. (cat. no. 310-04). Rocaglate CR-1-31B was synthesized as previously described (12 (link)) and other reagents were obtained from Sigma-Aldrich; Merck KGaA, unless otherwise stated.
Human GBC and paired normal gallbladder tissues were collected from 42 patients during gallbladder resection for GBC (22 males and 20 females) between July 2015 and June 2018 at the Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. The patients did not receive chemotherapy prior to surgery. The use of human GBC tissues was approved by Tongji Hospital Research Ethical Committee, and written informed consent was obtained from all the patients.

Cao Y., He Y., Yang L, & Luan Z. (2020). Targeting eIF4A using rocaglate CR-1-31B sensitizes gallbladder cancer cells to TRAIL-mediated apoptosis through the translational downregulation of c-FLIP. Oncology Reports, 45(1), 230-238.

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Primary Cell Culture Protocols

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Primary aortic smooth muscle cells (aSMCs) were purchased from Lonza (CC-2571) and maintained in Smooth Muscle Growth Medium (CC-3182; Lonza). Primary Human Renal Proximal Tubular Epithelial Cells (HRPTECs) were purchased from ScienCell (#4100) and maintained in Epithelial Cell Medium (#4101; ScienCell). Immortalized Human Kidney [48 (link)] (HK2) cells were maintained in RPMI-1640 medium with glutamine (61,870,010; ThermoFisher), supplemented with 10% (vol/vol) FCS (Biowest) and 100 UmL⁻¹ PS (Gibco). All cells were cultured at 37 °C in a humidified atmosphere containing 5% CO2. Primary cell lines were used between passages 3 and 8.

Louzao-Martinez L., van Dijk C.G., Xu Y.J., Korn A., Bekker N.J., Brouwhuis R., Nicese M.N., Demmers J.A., Goumans M.J., Masereeuw R., Duncker D.J., Verhaar M.C, & Cheng C. (2019). A proteome comparison between human fetal and mature renal extracellular matrix identifies EMILIN1 as a regulator of renal epithelial cell adhesion. Matrix Biology Plus, 4, 100011.

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Senescence Induction in Human Renal Epithelial Cells

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Human renal proximal tubular epithelial cells (HRPTEpiC) (ScienCell, Carlsbad, CA, USA) were grown in an epithelial cell medium (ScienCell, Carlsbad, CA, USA) containing epithelial cell growth supplement (ScienCell, Carlsbad, CA, USA), in a humidified atmosphere of 95% air and 5% CO₂ at 37 °C. HRPTEpiC was used at passages 8–10. Cells were plated at a density of 3 × 10⁵ cells/well in 6-well plates and incubated for 3 days. Fresh medium containing doxorubicin (DOXO) (Sigma, St. Louis, MO, USA) was added to cells and cultured for 24-h to induce cellular senescence. Similarly, HRPTEpiC was treated with fresh medium containing H₂O₂ (Sigma, St. Louis, MO, USA). Cells were harvested at the end of the treatment for further analysis.

Jin Y., Kim E.N., Lim J.H., Kim H.D., Ban T.H., Yang C.W., Park C.W, & Choi B.S. (2021). Role of Aberrantly Activated Lysophosphatidic Acid Receptor 1 Signaling Mediated Inflammation in Renal Aging. Cells, 10(10), 2580.

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Cell Culture Protocol for Renal Cell Lines

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ACHN, CAKI-1, 769-P and 786-O were obtained from ATCC (Va, USA) and used within twenty times of passages. Human renal proximal tubular epithelial cells (HPTEC) were acquired from ScienCell (ScienCell, CA, USA) and made use of within passage five. The ACHN and CAKI-1 were cultured in Dulbecco’s modified Eagle medium (Gibco, MA, USA). 769-P and 786-O were cultured in RPMI 1640 medium (Gibco, MA, USA) and HPTEC were cultured in Epithelial Cell Medium (ScienCell, CA, USA). All the mediums were supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin. Cells were seeded in a T25 flask or 6-well plate at 37 °C in a 5% CO₂ humidified incubator and collected for further experiments when reaching approximately 80–85% confluence.

Wang L.H., Cao B., Li Y.L, & Qiao B.P. (2023). Potential prognostic and therapeutic value of ANXA8 in renal cell carcinoma: based on the comprehensive analysis of annexins family. BMC Cancer, 23, 674.

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Morpholino-ASO Enhances SMN Protein Expression

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A previous study observed that morpholino-ASO was able to significantly increase the expression of SMN protein (16 (link)). Therefore, morpholino-ASO was purchased from Gene Tools, LLC, Philomath, OR, USA). The morpholino-ASO sequence was ATT CAC TTT CAT AAT GCT GG, targeting intronic splicing silencer N1 (ISS-N1) in SMN2 intron 7. SMA-01 and SMA-13 cell lines were adopted. The doses of ASO used were 0, 10, 20 and 40 pmol/well. Morpholino-ASO intervention was performed using an electroporator (BEX CO., LTD., Tokyo, Japan) and Opti-MEM (Gibco; Thermo Fisher Scientific, Inc.) was used as an electroporation medium, with a final volume of 30 µl/well. The parameters of electroporation were: Poration pulse (Pp) V, 150 V; Driving pulse (Pd) V, 20 V; Pd cycle, 10; Pp on, 10.0 msec; Pd on, 50.0 msec; Capacity (Capa), 1416.3 uF; Pp off, 10.0 msec and Pd off, 50.0 msec. Following electroporation, urine cells were seeded onto 12-well plates with 3×104 cells/well in epithelial cell medium (ScienCell Laboratories, Inc.) at 37°C for 6 h. After 6 h, the medium was switched to fresh epithelial cell medium (ScienCell Laboratories, Carlsbad, CA, USA). SMN protein was harvested 24, 48 and 72 h after seeding. All experiments were repeated at least three times.

Zhang Q.J., Lin X., Li J.J., Lu Y.Q., Guo X.X., Dong E.L., Zhao M., He J., Wang N, & Chen W.J. (2017). Application of urine cells in drug intervention for spinal muscular atrophy. Experimental and Therapeutic Medicine, 14(3), 1993-1998.

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Culturing Primary Human Nonpigmented Ciliary Epithelial Cells

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Primary HNPCE cells were purchased from ScienCell Research Laboratories (cat no. 6580, CA, USA). The cells were cultured in epithelial cell medium (ScienCell Research Laboratories, cat no. 4101, CA, USA) at 37 °C in an incubator with 5% CO₂ and 95% humidity according to the instruction of manufacturer. Originally, the cells were derived from clones obtained from a primary culture of human nonpigmented ciliary epithelium [12 (link)].

Zheng W., Ziemssen F., Suesskind D., Voykov B, & Schnichels S. (2023). TRPP2 is located in the primary cilia of human non-pigmented ciliary epithelial cells. Graefe's Archive for Clinical and Experimental Ophthalmology, 262(1), 93-102.

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Synergistic Effects of Sulforaphane and Gemcitabine in iCCA

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d,l-sulforaphane (1-isothiocyanate-4-methylsulphinylbutane, purity ≥ 98%) was purchased from Toronto Research Chemicals Inc. (Toronto, ON, Canada), and gemcitabine (2′-deoxy-2′,2′-difluorocytidine, purity ≥ 98%) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Two human iCCA cell lines, HuCCT-1 (cat: JCRB0425) and HuH28 (cat: JCRB0426) were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). These cells were cultured in RPMI-1640 (Nacalai Tesque, Inc., Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS) and 1% ampicillin/streptomycin. The primary human biliary epithelial cell line (HIBEpiC, cat: #5100) was purchased from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA). HIBEpiC cells were cultured in Epithelial Cell Medium (ScienCell Research Laboratories) supplemented with 2% FBS and 1% epithelial cell growth supplement (ScienCell Research Laboratories), and 1% ampicillin/streptomycin. The cells were grown at 37 °C in a 5% CO₂ atmosphere.

Tomooka F., Kaji K., Nishimura N., Kubo T., Iwai S., Shibamoto A., Suzuki J., Kitagawa K., Namisaki T., Akahane T., Mitoro A, & Yoshiji H. (2023). Sulforaphane Potentiates Gemcitabine-Mediated Anti-Cancer Effects against Intrahepatic Cholangiocarcinoma by Inhibiting HDAC Activity. Cells, 12(5), 687.

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Tissue-Engineered Vascular Graft Protocol

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Sodium dodecyl sulfate (SDS), Triton X-100 and trypsin were purchased from Sangon Biotech (Shanghai) Co., Ltd. PLGA (LA:GA = 85:15, ŋ = 3.1DL/g) was purchased from Corbion Trading (Shanghai) Co., Ltd. 2,2,2-trifluoroethanol (purity 99.5%) was purchased from Shanghai Aladdin Bio-Chem Technology Co., Ltd. GenElute™ Mammalian Genomic DNA Miniprep Kit and radio immunoprecipitation assay (RIPA) buffer were purchased from Sigma-Aldrich. Quant-iT PicoGreen dsDNA Assay Kit and 4′,6-Diamidine-2-phenylindole (DAPI) were purchased from Invitrogen. Epithelial cell medium and smooth muscle cell medium were purchased from Sciencell Research Laboratories, Inc. Cell Counting Kit-8 (CCK-8) was purchased from Dojindo. Hematoxylin and Eosin Staining Kit (H&E) and bicinchoninic acid (BCA) protein assay kit were purchased from Shanghai Beyotime Biotechnology Co., Ltd. Anti-Cytokeratin-14 (CK-14) antibody, anti-actin antibody and horseradish peroxidase (HRP)-conjugated secondary antibody were purchased from ABCam.

Qiu S., Liang L., Zou P, & Chen Q. (2021). Decellularized small intestine submucosa/polylactic-co-glycolic acid composite scaffold for potential application in hypopharyngeal and cervical esophageal tissue repair. Regenerative Biomaterials, 8(2), rbaa061.

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Primary RPE and Choroidal Endothelial Cells

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Human primary RPE cells (HRPEpiC) were purchased from ScienCell (Carlsbad, CA, USA) and maintained in epithelial cell medium (ScienCell). Rhesus macaque choroidal endothelial cells (RF/6A) were purchased from American Type Culture Collection (Manassas, VA, USA). RF/6A cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Invitrogen), 100 units/mL penicillin, and 100 µg/mL streptomycin (Gibco, Invitrogen,). RPE cells were cultured for two weeks prior to experiments, and experiments were conducted in epithelial cell medium. CECs were serum-starved in DMEM with 2% FBS overnight prior to experiments, and all RF/6A experiments were performed in DMEM with 0.5% FBS. Cells were incubated at 37 °C, 5% CO₂, 20.9% O₂, and 95% relative humidity. Sodium glycocholate (Sigma-Aldrich #G7132, St. Louis, MO, USA), sodium glycodeoxycholate (Calbiochem, Millipore #361311, Burlington, MA, USA), and sodium glycoursodeoxycholate (Toronto Research Chemicals #G679545, Toronto, ON, Canada) were dissolved in water to make 10 mM stock solutions for use in experimental assays.

Warden C, & Brantley MA J.r. (2021). Glycine-Conjugated Bile Acids Protect RPE Tight Junctions against Oxidative Stress and Inhibit Choroidal Endothelial Cell Angiogenesis In Vitro. Biomolecules, 11(5), 626.

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Isolation and Culture of HERS Cells and DPCs

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Primary HERS cells and DPCs were isolated from the first molar germ in the maxilla and mandible of 8-day-postnatal Sprague-Dawley (SD) rats (Figure S1), and the specific method was mentioned in the previous studies[26 (link),27 ]. After being sheared to fragments, the tissue was digested by type I collagenase and dispase (Sigma-Aldrich, USA) solution at 37°C for 10–20 min.
HERS cells were cultured in epithelial cell medium (ScienCell, USA) containing 1% epithelial cell growth supplement, 2% fetal bovine serum, and 1% penicillin/streptomycin solution. Mixed with mesenchymal cells, HERS cells in passage 0 were treated with 0.25% trypsin-EDTA (Gibco, USA) for 2–3 minutes, then purified HERS cells were obtained after 2–3 times of differential digestion. The adherent HERS cells were processed into cell suspensions under the treatment of TrypLE Express Enzyme (Thermo Scientific, USA) for 10–15 min. DPCs were cultured in alpha-minimum essential medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA) and 1% penicillin/streptomycin solution (Solarbio, CHN). All cells were cultured in cell incubator with 5% CO₂ at 37°C and medium was changed every 2 days.

Tang H., Bi F., Chen G., Zhang S., Huang Y., Chen J., Xie L., Qiao X, & Guo W. (2022). 3D-bioprinted Recombination Structure of Hertwig’s Epithelial Root Sheath Cells and Dental Papilla Cells for Alveolar Bone Regeneration. International Journal of Bioprinting, 8(3), 512.

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