The human prostate cancer cell line, PC-3, was purchased from the RIKEN Cell Bank (Osaka, Japan). Cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and Penicillin-Streptomycin Mixture (FUJIFILM Wako Pure Chemicals). All cells were cultured in a humidified atmosphere containing 5% CO2 at 37 °C. The PrimeSurface® system (Sumitomo Bakelite, Tokyo, Japan) was used for three-dimensional spheroid formation. Cell suspensions were seeded onto a PrimeSurface 96U plate at 104 cells/well, and then cultured for 7 days.
Penicillin streptomycin mixture
Manufactured by Fujifilm
Sourced in United States
Penicillin-streptomycin mixture is a common antibiotic solution used in cell culture applications. It is a combination of the antibiotics penicillin and streptomycin, which work together to inhibit the growth of a wide range of bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Primesurface system, by Sumitomo Bakelite (1 mentions)
Interleukin 2 (il 2), by Merck Group (1 mentions)
Rpmi 1640 medium, by Fujifilm (1 mentions)
Glass bottom culture dishes, by MatTek (1 mentions)
Alexa 488 conjugated anti mouse igg antibody, by Thermo Fisher Scientific (1 mentions)
Nucred live 647 readyprobes reagent, by Thermo Fisher Scientific (1 mentions)
α mem, by Merck Group (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Neurobasal medium, by Thermo Fisher Scientific (1 mentions)
Retinoic acid, by Merck Group (1 mentions)
Penicillin and streptomycin mixture, by Fujifilm (1 mentions)
Ultra low attachment culture dishes, by Corning (1 mentions)
α mem, by Fujifilm (1 mentions)
5 protocols using penicillin streptomycin mixture
1
3D Spheroid Culture of PC-3 Prostate Cancer Cells
2
Maintaining mCTLL-2 Cell Line
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The mCTLL-2 was supplied by the RIKEN BioResource Center (RIKEN BRC) (Tsukuba, Japan). Cells were maintained at 37 °C, in 5% CO2 with an RPMI 1640 medium (FUJIFILM Wako Pure Chemical) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA), a penicillin-streptomycin mixture (FUJIFILM Wako Pure Chemical), and 100 U/mL of IL-2 (Sigma-Aldrich).
3
Neurotransmitter Transport Assay Protocol
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Glass-bottom culture dishes were purchased from MatTek Corporation (Ashland, OR, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS) and penicillin/streptomycin mixture were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan), Biowest (Rue de la Caille, France) and Nacalai Tesque (Kyoto, Japan), respectively. An anti-DYKDDDDK tag mouse monoclonal antibody was purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan). An Alexa488-conjugated anti-mouse IgG antibody was purchased from Molecular Probe (Eugene, OR, USA). NucRed® Live 647 ReadyProbes® Reagent was purchased from Thermo Fischer Scientific (Tokyo, Japan); 4-phenylbutylate (4-PBA) and SKF-10047 were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan) and Tocris Bioscience (Bristol, UK), respectively; and [3H] 5-HT (370 GBq/mmol) was purchased from PerkinElmer (Waltham, MA, USA). The Neurotransmitter Transport Assay Kit was purchased from Molecular Device Corporation (Sunnyvale, CA, USA). All other chemicals were of analytical grade.
4
Overexpression of Human LRH1 in Cell Lines
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Human hepatocellular carcinoma cell lines HLF (JCRB0405) and JHH6 (JCRB1030) were purchased from JCRB cell bank (Osaka, Japan). Huh7.5.1 was gifted from Professor Matsuura, Osaka University. purchased from American Type Culture Collection. These cell lines were grown in Dulbecco’s Modified Eagle Medium (DMEM; D7777; Millipore Sigma, Burlington, MA, USA) with 10% fetal bovine serum (FBS; Millipore Sigma, Burlington, MA, USA) and 1% penicillin-streptomycin mixture (168-23191; FUJIFILM Wako Pure Chemical, Osaka, Japan). To overexpress human LRH1, the protein-coding region of hLRH1 was cloned into the NotI/BamHI site of the CSII-EF-MCS-IRES2-Venus plasmid (RDB04384; Riken, Ibaraki, Japan). The hLRH1S510A mutant was established by a PCR-based standard mutagenesis protocol. The plasmids were transiently transfected to 293T cells using Polyethylenimine Max (24765-1; Polysciences, Warrington, PA, USA).
5
Neuronal Differentiation of P19 Cells
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The P19 embryonal carcinoma cells (ATCC, Np.CRL-1825) were cultured with α-MEM (Sigma-Aldrich) and 10% fetal calf serum, GlutaMAX (Thermo Fisher Scientific), and penicillin-streptomycin mixture (Wako) (α-MEM/10% fetal calf serum) in a 5% CO 2 humidified atmosphere at 37°C. For neuronal differentiation, P19 cells were suspended in α-MEM/10% fetal calf serum medium containing 500 nM retinoic acid (Sigma-Aldrich) and were cultured on ultra-low attachment culture dishes (Corning, Lowell, MA). Four days after retinoic acid treatment, the cell aggregates were harvested and washed with PBS. Then, the cells were trypsinized, passed through a 70 μm nylon mesh (Falcon 352350; Becton Dickinson, Franklin Lakes, NJ), and plated onto a BioCort poly-D-lysine coated 12 well plate (Corning) at a density of 4.0 × 10 5 cells/well. Four hours after seeding, the medium was replaced with neurobasal medium (Thermo Fisher Scientific) supplemented with B-27 and penicillin and streptomycin mixture (Wako) (Neurobasal/B-27).
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