The human cervical cancer cell line HeLa was obtained from Keio University in Japan, and cultured in Ham's F12 medium (Wako Pure Chemical, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS; Gibco Life Technologies; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Wako Pure Chemical). Cells were incubated at 37°C in 5% CO2. To confirm the identity of the analyzed cell line, we performed short tandem repeat (STR) genotyping, which revealed correspondence with more than 80% of the markers tested. We found no mycoplasma in cells tested with MycoAlert mycoplasma detection kit (Lonza).
Ham s f12 medium
Manufactured by Fujifilm
Sourced in Japan
Ham's F12 medium is a cell culture medium commonly used in laboratory settings. It provides a balanced salt solution and essential nutrients to support the growth and maintenance of various cell types. The medium's core function is to facilitate the in vitro cultivation of cells, enabling researchers to study cellular processes and behavior.
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Lab products found in correlation
Rpmi 1640 medium, by Fujifilm (4 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (4 mentions)
Penicillin streptomycin, by Fujifilm (3 mentions)
Penicillin, by Fujifilm (3 mentions)
Dulbecco s modified eagle s medium, by Fujifilm (3 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (2 mentions)
Streptomycin, by Fujifilm (2 mentions)
Leiboviz s l 15 medium, by Fujifilm (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mem non essential amino acid, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Mycoalert mycoplasma detection kit, by Lonza (1 mentions)
N 2 hydroxyethylpiperazine n 2 ethane sulfonic acid hepes, by Thermo Fisher Scientific (1 mentions)
Mouse tpo, by Thermo Fisher Scientific (1 mentions)
Mouse scf, by Thermo Fisher Scientific (1 mentions)
Polyvinyl alcohol (pva), by Merck Group (1 mentions)
Hygromycin b, by Fujifilm (1 mentions)
Hypoxanthine aminopterin thymidine medium, by Merck Group (1 mentions)
30 protocols using ham s f12 medium
1
Culturing HeLa Cervical Cancer Cells
2
Expansion of mouse hematopoietic stem/progenitor cells
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C57BL/6-CD45.1 mouse BM cells, unfractionated or following magnetic c-Kit+ cell enrichment, were cultured using Ham’s F-12 medium (Wako), supplemented with 0.1% PVA (Sigma, Cat# P8136), 1% Insulin-Transferrin-Selenium-Ethanolamine (ITS-X) (100X) (Thermo Fisher Scientific), 1% Penicillin-Streptomycin-L-Glutamine Solution (100X) (Wako), N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES) (10 mM; Gibco), mouse TPO (100 ng/ml; PeproTech), and mouse SCF (10 ng/ml; PeproTech) for 28 days, incubated at 37 °C in a humidified 5% CO2 incubator. Medium was changed every other day15 (link),16 (link). Magnetic cell separation was performed using anti-mouse c-Kit MicroBeads (Miltenyi Biotech, Cat# 130-091-224) according to the manufacturer’s instructions. Cell culture using either commercially available pre-packaged HemEx-Type9A (NIPRO) or in-house prepared medium supplemented with 100 ng/mL mouse TPO and 10 ng/mL mouse SCF. Unfractionated whole BM cells were seeded at 2 × 106/mL onto 100 mm dish in 10 mL culture medium (day 0–14) or 60 mm dish in 4 mL culture medium (day 15–28). Magnetic column-enriched c-Kit+ BM cells were seeded at 1 × 106/well onto 48-well plates in 1 mL culture medium. For long-term cultures, complete medium changes were made every 2 days and cell cultures were passaged at a ratio of 1:2-3 when cells exhibited 80–90% confluency.
3
Culturing CHO and HEK293 cells with MAC4
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CHO cells containing MAC4 (CHO/MAC4) were cultured in Ham’s F12 medium (Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS) (Biowest, Vieux Bourg, France), 1% penicillin/streptomycin (Wako), and 800 µg/mL hygromycin B (Wako) (Takiguchi et al. 2012 (link); Narai et al. 2015 (link)). CHO/MAC4 cells containing a reconstructed hypoxanthine–guanine phosphoribosyl transferase (HPRT) gene and desired genes were selected in Ham’s F12 medium with 2% hypoxanthine-aminopterin-thymidine (HAT) medium (Sigma-Aldrich, St. Louis, MO, USA). HEK293 cells purchased from the American Type Culture Collection (catalog number CRL-157, Manassas, VA, USA) were cultured in Eagle’s minimum essential medium (MEM) (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% FBS, 1% MEM non-essential amino acids (Thermo Fisher, Waltham, MA, USA), and 1% l -glutamine (Thermo Fisher). HEK293 cells containing MAC4 with the desired genes were selected in Eagle’s MEM containing 200 µg/mL hygromycin B.
4
Cultivation of Diverse Cell Lines
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Human fibrosarcoma (HT1080) cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Sigma) plus 10% fetal bovine serum (FBS). HFL-1 cells (RCB0521, RIKEN, Tsukuba, Japan) were grown in Ham’s F12 medium (Wako Pure Chemical Industries) supplemented with 15% FBS. Hprt-deficient Chinese hamster ovary (CHO) cells (JCRB0218, JCRB Cell Bank, Japan) containing the HAC vector were cultured in Ham’s F-12 medium supplemented with 10% FBS and 8 μg/ml Blasticidin S (Bsd, Funakoshi). HFL-1 cells introduced the HAC vector for reprogramming (designated as iHAC) were maintained on mitomycin-C (Kyowa Hakko Kirin)-treated SNL (STO) feeder cells (SANGER Institute, Cambridge, UK) in hES cell maintenance medium, which consisted of a 1:1 DMEM and Ham’s F-12 (Sigma) supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 1% MEM non-essential amino acids, 0.1 mM 2-mercaptoethanol (Gibco), 4 ng/ml human basic fibroblast growth factor (Wako Pure Chemical Industries) and 20% Knockout serum replacement (Gibco).
5
Maintenance of Human and Canine Prostate Cancer Cell Lines
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The human 293 T and PC3 cell lines were provided by the American Type Culture Collection (ATCC, Rockville, MD). The canine prostate cancer CHP-1 cell line was established from a prostate mass collected immediately following surgery of a tumour-bearing, 10-year-old, castrated male Jack Russell Terrier breed dog in our university [5 (link)]. The 293 T and the CHP-1 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Wako, Osaka, Japan) and PC3 cells were maintained in Ham’s F12 medium (Wako) supplemented with 10% foetal bovine serum (FBS), penicillin (50 IU/mL), and streptomycin (50 μg/mL) under a humidified atmosphere with 5% CO2 at 37 °C.
6
Cancer Cell Line Culture Protocols
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The esophageal cancer cell line KYSE30 (94072011; European Collection of Authenticated Cell Cultures, Salisbury, UK) was cultured in a 1:1 mixture of Ham’s F12 medium (Wako Pure Chemical Industries, Tokyo, Japan) and RPMI 1640 medium (Wako Pure Chemical Industries) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Biosera, Nuaillé, France) and 1% ampicillin and streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). The human colon cancer cell line HCT116 (CCL-247; American Type Culture Collection, Manassas, VA, USA) was cultured in RPMI 1640 medium (Wako Pure Chemical Industries) supplemented with 10% FBS and 1% ampicillin and streptomycin (Thermo Fisher Scientific). Murine colon adenocarcinoma cell line MC38 (ENH204-FP; Kerafast, Boston, MA, USA) was cultured in Dulbecco’s modified Eagle medium with high glucose (Wako Pure Chemical Industries) supplemented with 10% FBS, 1 mM sodium pyruvate (Thermo Fisher Scientific), 100 μM non-essential amino acids (Thermo Fisher Scientific), 50 μg/mL gentamicin (Wako Pure Chemical Industries), 10 μM 4-(2-hydroxyethyl)-1-piperazineëthanesulfonic acid (Thermo Fisher Scientific), and 1% penicillin-streptomycin-amphotericin B (Wako Pure Chemical Industries). Cells were cultured in an atmosphere of 5% carbon dioxide (CO2) at 37°C. All experiments using cells in this study were performed for fewer than 20 passages after thawing.
7
Cell Culture and Transfection of B16 and HMV-II Cell Lines
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B16 (RRID: CVCL_F936) and HMV-II (RRID: CVCL_1282) cells were purchased from RIKEN BRC (Ibaraki, Japan). B16 cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Fujifilm Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (fetal bovine serum [FBS]; Nichirei – lot number S.18M00C) [28 (link)]. HMVII cells were maintained in Ham’s F12 medium (Fujifilm Wako), supplemented with 10% FBS. HMV-II cells were authenticated by STR (or SNP) profiling by RIKEN BRC. B16 and HMV-II cells were cultured in mycoplasma-free conditions. Cells were transfected using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA), according to the manufacturer’s instructions.
8
Isolation and Culture of KMCs
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KMCs from trap:GFP animals were resuspended in medium containing 40% Leiboviz’s L-15 medium (Wako), 32% Dulbecco’s modified Eagle’s medium (Wako), 12% Ham’s F12 medium (Wako), 8% FBS, 2 mM l -glutamine (Wako), 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Sigma), 100 U penicillin (Wako), and 100 μg/mL streptomycin (Wako). Approximately 6 × 104 of KMCs were plated on a 96-well plate coated with fibronectin (Corning) and incubated for 3 h at 30 °C, 5% CO2. Subsequently, ~2000 of sorted cells or EVs were plated in the wells (four wells for each fraction) and incubated for additional 2 days at the same condition described above. The number of GFP+ cells was counted using an EVOS FL Cell Imaging System (Thermo Fisher Scientific). For counting the number of nuclei, Hoe was directly added into the medium at a concentration of 5 μg/mL. For counting the total number of cells in each well, cells were treated with 0.25% trypsin–1 mM EDTA and harvested by pipetting. The total number of cells was counted using a hemocytometer (Funakoshi).
9
Cell Culture of Human Cell Lines
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The human embryonic kidney cell line 293T and the human prostate cancer cell lines PC3 and LNCaP were provided by American Type Culture Collection (Rockville, MD). The normal human foreskin fibroblast cell line Hs68 was provided by JCRB Cell Bank (Osaka, Japan). The 293T, LNCaP and Hs68 cells were maintained in Dulbecco's modified Eagle's medium (DMEM) (Wako, Osaka, Japan), and PC3 cells were maintained in Ham's F12 medium (Wako) supplemented with 10% fetal bovine serum (FBS), penicillin (50 IU/ml) and streptomycin (50 μg/ml) under a humidified atmosphere of 5% CO2 at 37°C.
10
Culturing Pv11 and CHO Cells
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Pv11 cells and CHO cells were grown as previously described (9 (link), 14 (link), 82 (link)). Pv11 cells were cultured in IPL-41 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 2.6 g/L tryptose phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum (FBS) and 0.05% (v/v) of Antibiotic-Antimycotic Solution (10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL; Sigma–Aldrich, St. Louis, MO), designated hereafter as complete IPL-41 medium. A CHO-K1 derivative cell line, Flip-InTM-CHO cells (purchased from Thermo Fisher Scientific), was cultured in Ham’s F-12 medium (FUJIFILM Wako, Osaka, Japan) containing 10% (v/v) FBS, 100 units/mL penicillin and 100 μg/mL streptomycin (penicillin-streptomycin mixture; Sigma–Aldrich) at 37 °C, 5% CO2, and 95% relative humidity.
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