Madin-Darby canine kidney (MDCK) cells (ATCC) and RAW264.7 (ATCC) cells, murine macrophages, were cultured on 35 mm diameter six well cell culture plates (SPL Life Sciences) in minimum essential medium (MEM) (Corning) with 5% fetal bovine serum (FBS; MP Biomedicals) and 100 unit/ml streptomycin/penicillin (S/P; Welgene Inc.) at 37°C in a humidified atmosphere containing 5% CO2.
Penicillin streptomycin p s
Manufactured by Welgene
Sourced in United States
Penicillin-streptomycin (P/S) is a commonly used antibiotic solution that contains a combination of the antibiotics penicillin and streptomycin. It is a sterile liquid designed for cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Welgene (10 mentions)
Dulbecco s modified eagle s medium dmem, by Welgene (2 mentions)
Zinc nitrate hexahydrate, by Merck Group (2 mentions)
Tween 80, by Samchun (2 mentions)
Absolute alcohol, by Samchun (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Welgene (2 mentions)
Phosphate buffered saline (pbs), by Welgene (2 mentions)
Cell culture plate, by SPL Life Sciences (1 mentions)
Minimum essential medium (mem), by Corning (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Atcc tib 71, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Gold 3 chloride trihydrate, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Junsei (1 mentions)
Methacrylic anhydride, by Merck Group (1 mentions)
Pegdma, by Polysciences (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline pbs, by Biosesang (1 mentions)
Trypsin ethylenediaminetetraacetic acid, by Thermo Fisher Scientific (1 mentions)
22 protocols using penicillin streptomycin p s
1
MDCK and RAW264.7 Cell Culture
2
Immunofluorescence of Primary Cilia in MDCK Cells
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Madin-Darby canine kidney cells (MDCK, American Type Culture Collection) were cultured in MEM with 5% FBS (Mediatech Inc.) with streptomycin/penicillin (S/P) 100 unit/ml (WelGENE Inc., Daegu, Korea). For immunofluorescence assays of primary cilia, cells were cultured on coverslips. After becoming confluent, cells were treated with hydrogen peroxide (Sigma, St. Louis, MO), MnTMPyP, or vehicle for the indicated times and conditions. Cells were fixed with 4% paraformaldehyde and processed for immunofluorescence or lysed for Western blot analysis.
3
Murine Kidney Proximal Tubule Cell Proliferation
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Murine proximal tubular epithelial cells (mProx24) derived from C57BL/6J adult mouse kidney as described previously35 (link) were gifted by Dr. Kwon HM (UNIST, Ulsan, Korea). RAW 264.7 cells, a murine macrophage/monocyte were purchased from ATCC (ATCC® TIB-71™). mProx24 and RAW264.7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 15% fetal bovine serum (FBS) (Mediatech Inc., Manassas, VA, USA) with 100 u/ml streptomycin/penicillin (S/P) (WelGENE Inc., Daegu, Korea). mProx24 and RAW264.7 cells were seeded on 0.1% gelatin-coated culture dishes and normal culture dishes (Falcon, Oxnard, CA, USA), respectively. To determine mitotic cells, cells were fixed (4% paraformaldehyde) and then subjected to immunofluorescence- staining to evaluate mitotic spindle using anti-α-tubulin antibody. DAPI stain was used to visualize the nucleus. Proliferation of cells was evaluated mitotic spindle-stained by α-tubulin antibody and dividing nucleus by DAPI. Number of cells and mitotic cells were counted within 10 fields per slide.
4
Synthesis and Characterization of Gold Nanoparticles
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Gold(III) chloride trihydrate
(HAuCl4·3H2O, 99.9% trace metal basis),
agarose, and sodium chloride (NaCl) were purchased from Sigma-Aldrich
(USA) and were used without further purification. TEMPO-CNCs were
received from the Cellulose Lab (Canada) and used as obtained. Sodium
hydroxide (NaOH) was purchased from JUNSEI (Japan). The ssDNA probe
and target and non-target DNA were received from BIONEER Inc. (Daejeon,
Republic of Korea). Dulbecco’s modified Eagle’s medium
(DMEM), 10% fetal bovine serum (FBS), and antibiotics Penicillin–Streptomycin
(P/S) were purchased from Welgene Inc. (Republic of Korea). A WST-1
dye (EZ-Cytox cell viability assay kit) was purchased from DoGenBio
Co., Ltd. (Seoul, Republic of Korea).
(HAuCl4·3H2O, 99.9% trace metal basis),
agarose, and sodium chloride (NaCl) were purchased from Sigma-Aldrich
(USA) and were used without further purification. TEMPO-CNCs were
received from the Cellulose Lab (Canada) and used as obtained. Sodium
hydroxide (NaOH) was purchased from JUNSEI (Japan). The ssDNA probe
and target and non-target DNA were received from BIONEER Inc. (Daejeon,
Republic of Korea). Dulbecco’s modified Eagle’s medium
(DMEM), 10% fetal bovine serum (FBS), and antibiotics Penicillin–Streptomycin
(P/S) were purchased from Welgene Inc. (Republic of Korea). A WST-1
dye (EZ-Cytox cell viability assay kit) was purchased from DoGenBio
Co., Ltd. (Seoul, Republic of Korea).
5
Hydrogel Synthesis and Characterization
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PEGDMA (MW: 1000) was purchased from Polysciences (Warrington, PA, USA). Edible pigments were purchased from Lgreentech (Daejeon, Gyeonggi, South Korea). Penicillin/streptomycin (P/S) and phosphate buffered saline (PBS, pH 7.4) were purchased from WelGene (Daegu, Gyeongbuk, South Korea). Gelatin (Type A, 300 bloom from porcine skin), methacrylic anhydride (MA), and LAP (≥95%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other chemical agents used in this study were of analytical grade.
6
Culturing Human Foreskin Fibroblasts
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Human foreskin fibroblasts were obtained from Millipore (Catalog no. SCC058). Cells were maintained as per the protocol in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA), 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA), and 1% penicillin-streptomycin (P/S) (Welgene, Inc., Gyeongsan, Korea).
7
Isolation and Culture of Human Amniotic Epithelial Stem Cells
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Amniotic membranes were obtained from the placentas of pregnant women who underwent elective cesarean sections. The amniotic membranes were washed several times with phosphate-buffered saline (PBS; Biosesang, Seongnam, Korea) to remove all the blood [19 ]. To isolate hAESCs, amniotic membranes underwent digestion twice with 0.05% trypsin-ethylenediaminetetraacetic acid (Invitrogen, CA, USA) for 30 min at 37°C. Isolated hAESCs were cultured in 100 mm diameter cell culture dishes with α-minimal essential medium (α-MEM; Welgene, Daegu, Korea) supplemented with 10% fetal bovine serum (FBS; Welgene), 1% penicillin-streptomycin (P/S; Welgene), and 10 ng/mL epidermal growth factor (EGF; R&D Systems, Minneapolis, MN, USA) [16 (link), 20 (link)]. All the procedures described below were performed under sterile conditions (Fig. 1a ). This study was approved by the Institutional Review Board of the Korea University Anam Hospital, Seoul, Korea (no. 2009AN0036). All patients who participated in this study provided their written informed consent prior to the commencement of the study.
8
Hypoxic culture of human HCC cell lines
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Human HCC cell lines, Hep3B, SNU449, and Huh7 cells were obtained from the Korean Cell line Bank (Seoul, Republic of Korea). Hep3B and SNU449 cells were grown in Roswell Park Memorial Institute-1640 medium (Welgene, Daegu, Republic of Korea) supplemented with 10% fetal bovine serum (FBS) (Welgene) and 1% penicillin streptomycin (PS) (Welgene). Huh7 cells were cultured in Dulbecco’s modified Eagle’s medium containing 10% FBS and 1% PS. For hypoxic incubation, cells were incubated in a hypoxic incubator (New Brunswick Scientific, Edison, NJ, USA) with a humidified environment consisting of 1% O2, 5% CO2, and 94% N2.
9
Adipogenic Differentiation of 3T3-L1 Preadipocytes
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3T3-L1 preadipocytes were maintained in Dulbecco’s Modified Eagle Medium (DMEM) (Welgene, Seoul, Korea) supplemented with 10% bovine calf serum (BCS) (Welgene, Seoul, Korea)and 1% penicillin/streptomycin (P/S) (Welgene, Seoul, Korea). To induce adipocyte differentiation, 3T3-L1 cells were incubated in adipogenic media consisting of DMEM, 10% fetal bovine serum (FBS) (Welgene, Seoul, Korea), 1% P/S, 10 μg/mL insulin (Sigma-Aldrich, St.Louis, MO, USA), 0.5 mM 3-isobuyl-1-methylxanthine (IBMX) (Sigma-Aldrich, St.Louis, MO, USA) and 1 μM dexamethasone (Sigma-Aldrich, St.Louis, MO, USA). Then, media were changed every 2 days with DMEM containing 10% FBS, 1% P/S, and 10 μg/mL insulin. In our experiments, to assess the effects of MAOC on adipogenesis, the cells were treated with MAOC during adipogenesis. Eight days after initiating adipogenesis, the cells were prepared for further experiments, including RT-qPCR and Western blotting.
10
Keratinocyte and Lung Cancer Cell Assays
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Zinc nitrate hexahydrate (>98.0%), indole-3-Carbinol (Sigma Aldrich, St. Louis, MO, USA), sodium hydroxide (>98.0%). All media was supplied from Difco, MB Cell (Gangnam-gu, Seoul, Korea). Absolute alcohol was supplied by Samchun Pure Chemical Co. Ltd. (Gyeonggi-do, Korea). Olive oil and Tween 80 were obtained from Samchun Pure Chemical Co. Ltd. The human keratinocyte cell line (HaCaT) and lung cancer cell line (A549) used in this study were obtained from the Korean cell line bank (Seoul, South Korea). RPMI 1640 (Roswell Park Memorial Institute Medium) culture medium was purchased from GenDEPOT Inc. (Barker, TX, USA). Dulbecco’s modified eagle’s medium (DMEM) (Gibson-BRL, Grand Island, NY, USA) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (p/s) (WElGENE Inc., Daegu, Korea) was also used for experiments.
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