Human amyloid β-protein (Aβ, Human, 1–42) was purchased from Peptide Institute (Osaka, Japan). DMEM Ham’s F-12 medium and all-trans retinoic acid (ATRA) were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Penicillin G sodium, streptomycin sulfate, amphotericin B, fetal bovine serum (FBS), Cur, and FA were obtained from Thermo Fisher Scientific K.K. (Waltham, MA, USA). Cur and FA were dissolved in dimethyl sulfoxide (DMSO) and then disbanded in medium to achieve a final concentration of DMSO to 0.1%. The other chemicals used in this experiment were the purest commercially available.
Dmem ham s f 12 medium
Manufactured by Fujifilm
Sourced in Japan
DMEM/Ham's F-12 medium is a cell culture medium commonly used for the in vitro cultivation of a variety of cell types, including mammalian cells. It is a mixture of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 nutrient mixture, providing a balanced formulation of essential nutrients, vitamins, and growth factors to support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Rpmi 1640 medium, by Fujifilm (4 mentions)
Dmem medium, by Fujifilm (2 mentions)
Penicillin streptomycin, by Fujifilm (2 mentions)
Rpmi 1640, by Fujifilm (2 mentions)
Penicillin, by Nacalai Tesque (2 mentions)
Streptomycin, by Nacalai Tesque (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (2 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Streptomycin sulfate, by Thermo Fisher Scientific (1 mentions)
Penicillin g sodium, by Thermo Fisher Scientific (1 mentions)
Dnase 1, by Roche (1 mentions)
Gfr matrigel, by Corning (1 mentions)
Stempro hesc sfm, by Thermo Fisher Scientific (1 mentions)
Liberase dh, by Roche (1 mentions)
Phg0311, by Thermo Fisher Scientific (1 mentions)
Nicotinamide, by Merck Group (1 mentions)
R spondin 2, by R&D Systems (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
23 protocols using dmem ham s f 12 medium
1
Amyloid β-Protein Inhibition by Curcumin and Ferulic Acid
2
Preparation and Xenograft of CTOS Lines
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For both surgical specimens and xenografts, CTOS were prepared as previously described with slight modifications.8 Briefly, tumors were mechanically minced and incubated for 90 minutes at 37°C with continuous stirring, in DMEM/Ham's F12 medium (Wako Pure Chemical Industries, Osaka, Japan) containing Liberase DH (Roche, Basel, Switzerland) at a final concentration of .28 units/mL. DNase I (Roche) was added at 10 μg/mL, followed by an additional 15 minutes of incubation. The digestion solution was serially strained using mesh filters of 500, 250, 100 and 40 μm (BD Falcon, Franklin Lakes, NJ, USA). Fractions were recovered between 100‐250 μm (Fr.100‐250) and 40‐100 μm (Fr.40‐100). Fr.40‐100 samples were cultured for 24‐48 hours at 37°C under 5% CO2, 20% O2, in STEMPRO hESC SFM (Invitrogen, Carlsbad, CA, USA) to form CTOS. Fr.100‐250 samples were mechanically disrupted by raising and lowering the plunger several times using a 27‐gauge needle, and then cultured as described for Fr.40‐100. To generate xenograft tumors of CTOS lines, freeze‐stocked CTOS12 were thawed, and cultured for 7 days in STEMPRO hESC SFM. We then suspended 2000 CTOS in a 1:1 mixture of medium and Matrigel GFR (Corning, Corning, NY, USA), and subcutaneously injected 1000 CTOS each into 2 sites of dorsal skin in NOD/scid mice.
3
Culture Media for Fallopian Tube Epithelial Cells
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We defined three types of culture medium for FTECs at different stages. The “basal medium” used DMEM/Ham’s F-12 medium (042-30795, Wako, Chuo, Osaka, Japan) supplemented with 1% GlutaMAX (35050061, Thermo, Waltham, MA, USA), 2% B27 (17504-044, Thermo), 1 mM nicotinamide (72340, Sigma-Aldrich, St. Louis, MO, USA), 0.5 µM transforming growth factor beta (TGFβ) receptor kinase inhibitor IV (SB431542, Wako), and 10 ng/mL human epidermal growth factor (EGF; PHG0311, Thermo). This medium was used as the essential medium for the short-term culture of primary FTECs to maintain an undifferentiated status in ALI culture. The “expansion medium” was rich in growth factors that support the proliferation of FTECs and maintained the stemness during expansion. It was based on basal medium supplemented with 5 µM Rho-associated coiled-coil containing kinase (ROCK) inhibitor (030-24021, Wako), 100 ng/mL human fibroblast growth factor (FGF, 100-26, Peprotech, Rocky Hill, NJ, USA), 100 ng/mL human Noggin (120-10C, Peprotech), 50 ng/mL Wnt-3a (5036-WN-010, R&D systems, Minneapolis, MN, USA), and 125 ng/mL R-Spondin 2 (3266-RS-025, R&D systems). Finally, the basal medium with 2 ng/mL β-estradiol (E4389, Sigma-Aldrich) was used as “differentiation medium” for inducing differentiation in ALI culture.
4
Ovarian Cancer Cell Line Cultivation and 5-Aza-2'-Deoxycytidine Treatment
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The serous ovarian cancer cell lines JHOS‐2, JHOS‐4 and NIH‐OVCAR3 were obtained from RIKEN BioResource Center (Tsukuba, Japan). WI‐38 and SKOV3 (epithelial ovarian cancer) were obtained from ATCC (Manassas, VA, USA). Although these cell lines were not authenticated, relatively low passage number cells were obtained. JHOS‐2 and JHOS‐4 were maintained in DMEM/Ham's F‐12 medium (Wako, Osaka, Japan), NIH‐OVCAR3 and SKOV3 were maintained in RPMI‐1640 medium (Wako), and WI‐38 was maintained in DMEM medium (Wako). All cell lines were cultured in medium containing 5% FBS (Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin‐streptomycin (Wako) at 37°C in a humidified incubator with 5% CO2. For 5‐aza‐2′‐deoxycytidine (DAC, Sigma‐Aldrich, St Louis, MO, USA) treatment, cells were treated with 500 nmol/L DAC for 3 days. Medium containing DAC was replaced every day. DNA and RNA were extracted on the 7th day following the treatment.
5
Canine Melanoma Cell Lines Characterization
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Five canine melanoma cell lines (CMM1, CMM2, CMM8, CMM11, and KMeC) were used [38 (link),39 (link),40 (link),41 (link)]. CMM1, CMM2, and KMeC were maintained in RPMI-1640 medium (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS; Cosmo Bio Co., Tokyo, Japan) and 50 mg/L gentamicin sulfate (Sigma Chemical Co., St Louis, MO, USA). CMM8 and CMM11 were maintained in D-MEM/Ham’s F-12 medium (FUJIFILM Wako Pure Chemical Corporation) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 250 ng/mL amphotericin B suspension (PSA; FUJIFILM Wako Pure Chemical Corporation). Chinese hamster ovary (CHO)-K1 and canine PDPN (dPDPN)-expressing CHO-K1 with N-terminal PA tag (CHO/dPDPN) cell lines were also used [15 (link)]. CHO-K1 and CHO/dPDPN were cultured in RPMI-1640 medium supplemented with 10% FBS and PSA. All cell lines were incubated at 37 °C in a humid atmosphere with 5% CO2. These conditions were used in the following experiments, unless otherwise stated.
6
Cell Culture Protocols for Diverse Cell Lines
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Nineteen cell lines were used in this study and specific details of each cell line are summarized in Table S2 . Cells were cultured in RPMI-1640 (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Thermo Fisher Scientific Inc., Waltham, MA, USA) and 5 mg/L gentamicin (Sigma-Aldrich Corp., St. Louis, MO, USA) or in DMEM/Ham’s F-12 medium (FUJIFILM Wako Pure Chemical Corporation) with 10% FBS and 100 U/mL penicillin, 100 µg/mL streptomycin (FUJIFILM Wako Pure Chemical Corporation), depending on the cell line (Table S2 ). Cells were maintained at 37 °C in a humidified atmosphere with 5% CO2.
7
Co-culture of Renal Cell Lines
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Human renal proximal tubule epithelial cells (HK-2 cells) and rat renal interstitial fibroblast (NRK-49F cells) were cultured in DMEM/Ham’s F-12 medium (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) with 10% FBS (Capricorn scientific, Ebsdorfergrund, Germany) and 1% antibiotic/antifungal mixture solution (100 U/mL penicillin, 100 μg/mL streptomycin, 0.25 μg/mL amphotericin B; Nacalai Tesque, Kyoto, Japan) at 37 °C in an atmosphere containing 5% CO2. Human monocytes (THP-1 cells) were cultured in RPMI 1640 (Gibco, Grand Island, NY, USA) with 10% FBS and 1% antibiotic/antifungal mixture solution. THP-1 cells were seeded and differentiated for 48 h by adding 50 nM phorbol 12-myristate 13-acetate (Sigma-Aldrich, St Louis, MO, USA). These cells were seeded in 6-well plates in appropriate cell numbers and used for the experiments (see legends for Figures).
8
Culturing hMSC-ATs using MSCGM-CD™ Medium
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The MSCGM-CD™ Mesencymal Stem Cell Growth Medium BulletKit™ was obtained from Lonza
(Basel, Switzerland). hMSC-ATs (46-year-old Caucasian female) (PromoCell, Heidelberg,
Germany) were cultured. Fetal bovine serum (FBS) was obtained from BioWest (Nuaille,
France). D-MEM/Ham’s F-12 medium was obtained from Wako (Osaka, Japan). Plastic dishes
were obtained from TPP (Trasadingen, Switzerland). All other materials used were of the
highest commercial grade.
(Basel, Switzerland). hMSC-ATs (46-year-old Caucasian female) (PromoCell, Heidelberg,
Germany) were cultured. Fetal bovine serum (FBS) was obtained from BioWest (Nuaille,
France). D-MEM/Ham’s F-12 medium was obtained from Wako (Osaka, Japan). Plastic dishes
were obtained from TPP (Trasadingen, Switzerland). All other materials used were of the
highest commercial grade.
9
Maintenance of Human Pluripotent Stem Cells
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Human ESCs (khES-2 strain obtained from Kyoto University) were maintained in an undifferentiated state as described previously (16) (link). Briefly, human ESCs were co-cultured with mouse embryonic fibroblasts (MEFs) in human ESC medium composed of 80% D-MEM/Ham’s F-12 medium (1/1 ratio; Wako Pure Chemical Industries, Osaka, Japan), 20% Knockout Serum Replacement (KSR; Invitrogen, Carlsbad, CA, USA), 0.1 mM nonessential amino acids (Sigma-Aldrich, St. Louis, MO, USA), 2 mM L-glutamine (Sigma-Aldrich), 0.1 mM β-mercaptoethanol (Sigma-Aldrich), and 4 ng/mL basic fibroblast growth factor (Wako).
Human iPSCs (253G1 strain obtained from Center for iPSC Research and Application, Kyoto University) were maintained under feeder-free conditions in Essential 8® medium (Invitrogen) on culture dishes coated with 0.5 μg/cm2 iMatrix-511 (Nippi, Tokyo, Japan).
Human iPSCs (253G1 strain obtained from Center for iPSC Research and Application, Kyoto University) were maintained under feeder-free conditions in Essential 8® medium (Invitrogen) on culture dishes coated with 0.5 μg/cm2 iMatrix-511 (Nippi, Tokyo, Japan).
10
Establishment of OSCC and Leukemia Cell Lines
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We used SCC25 TSCC and THP-1 human acute monocytic leukemia cell lines in this study. SCC25 is a common cell line for the OSCC research (24 –28 (link)), and many researchers have used 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated THP-1 cells as macrophage-like cells (19 (link), 29 (link)–32 (link)). Both cell lines were obtained from the American Type Culture Collection (Manassas, VA). SCC25 cells were routinely incubated in D-MEM/Ham’s F-12 medium (Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO) and 1% antibiotic–antimycotic (Invitrogen, Carlsbad, CA). THP-1 cells were incubated in RPMI-1640 (Wako) supplemented with 10% FBS (Sigma-Aldrich) and 1% antibiotic–antimycotic (Invitrogen). To induce macrophage-like differentiation, we treated THP-1 cells with 200 nM TPA (Cell Signaling Technology, Danvers, MA) as described previously (21 (link), 33 (link)). Before performing all experiments with macrophage-like cells, we aspirated the medium and washed the wells using PBS at least three times to remove TPA sufficiently from the well.
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