Human hair germinal matrix cells (HHGMCs; Catalog#2410) were purchased from ScienCell Research Laboratories (San Diego, CA, USA). The cells were cultured in a complete growth medium (mesenchymal stem cell medium (MSCM), supplemented with 5% fetal bovine serum (FBS), 1% stem cell growth supplement (MSCGS), and 1% penicillin/streptomycin solution (P/S) (all from ScienCell, San Diego, CA, USA) and incubated at 37 °C in a 5% CO2 humidified atmosphere. The medium was changed every 1–2 days.
Penicillin streptomycin solution p s
Manufactured by ScienCell
Sourced in United States
Penicillin/streptomycin solution (P/S) is a sterile, liquid culture supplement containing penicillin and streptomycin antibiotics. It is commonly used in cell culture applications to prevent bacterial contamination.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by American Type Culture Collection (4 mentions)
2 mercaptoethanol, by Merck Group (4 mentions)
Hypoxanthine, by American Type Culture Collection (4 mentions)
Thymidine, by American Type Culture Collection (4 mentions)
Fetal bovine serum (fbs), by ScienCell (4 mentions)
Sodium bicarbonate, by American Type Culture Collection (3 mentions)
Huvecs, by ScienCell (3 mentions)
Endothelial cell growth supplement ecgs, by ScienCell (2 mentions)
Endothelial cell medium ecm, by MP Biomedicals (2 mentions)
First passage normal hok, by ScienCell (2 mentions)
Oral keratinocyte medium, by ScienCell (2 mentions)
Huvecs, by Merck Group (1 mentions)
Iscove modified dulbecco media (imdm), by American Type Culture Collection (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Insulin, by Merck Group (1 mentions)
Padds, by ScienCell (1 mentions)
Hpa s, by ScienCell (1 mentions)
Astrocyte growth supplement, by ScienCell (1 mentions)
Ln229, by American Type Culture Collection (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
22 protocols using penicillin streptomycin solution p s
1
Culturing Human Hair Germinal Matrix Cells
2
HUVEC Cell Culture Protocol
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HUVECs were purchased from Sigma-Aldrich (St. Louis, MO). The cells were cultured in endothelial cell medium (ECM) supplemented with 5% (v/v) fetal bovine serum (FBS), 1% (v/v) endothelial cell growth supplement (ECGS) and 1% (v/v) penicillin/streptomycin solution (P/S) (ScienCell Research Laboratories, Inc., Cat No. 1001), at 37°C in a humidified atmosphere with 5% CO2. The medium was changed every other day unless specified.
3
Culturing SC Monocyte/Macrophage Cell Line
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All experiments were conducted on the commercially available monocyte/macrophage peripheral blood cell line SC (ATCC CRL-9855) (ATCC; Manassas, VA, USA). The cell line was maintained under standard conditions, according to the guidelines provided by the vendors (37 °C, 5% CO2, 95% humidity). The SC cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM; ATCC; Manassas, VA, USA) with 4 mM L-glutamine and 1.5 g/L sodium bicarbonate, which was supplemented with 0.05 mM 2-mercaptoethanol (Sigma-Aldrich Corp., St. Louis, MO, USA), 0.1 mM hypoxanthine, 0.016 mM thymidine (90%) (ATCC; Manassas, VA, USA), 10% fetal bovine serum (ATCC; Manassas, VA, USA), and 1% penicillin/streptomycin solution (P/S) (ScienCell Research Laboratories, San Diego ad, CA, USA).
4
Adipogenic Differentiation of Human and Mouse Preadipocytes
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Human preadipocyte (HPA-s) was purchased from ScienCell Research Laboratories (Cat. No. 7220; Carlsbad, CA, USA,) and mouse fibroblast 3T3-L cell lines was obtained from ATCC (Cat. No. ATCC® CL-173™; Manassas, USA,) and cultured in corresponding full medium in a humidified incubator at 37°C with 5% CO2.
Adipogenic differentiation of HPA-s was carried out according to previous methods14 (link). HPA-s were cultured for 3 days followed by stimulating with differentiation medium for 5 days. The differentiation medium consists of 500 ml of PADM basal medium (Cat. No. 7221, ScienCell), 25 ml of fetal bovine serum (FBS, Cat. No. 0025, ScienCell), 5 ml of preadipocyte differentiation supplement (PAdDS, Cat. No. 7232, ScienCell), 5 ml of penicillin/streptomycin solution (P/S, Cat. No. 0503, ScienCell). The medium was replaced every two days.
3T3-L1 preadipocytes were cultured for 2 days, then 3T3-L1 cells were stimulated with differentiation medium for 3 days. The differentiation medium consists of DMEM (Cat. No.11965092, ThermoFisher) with 10% FBS and MDI (0.5 mM 1-methyl-3-isobutylxanthine (Cat. No. I5879, Sigma-Aldrich), 1 μM dexamethasone (Cat. No.D4902, Sigma-Aldrich), and 10 μg/mL insulin, Cat. No. 12585014, Sigma-Aldrich). Cells were cultured with DMEM containing 10% FBS and 10 μg/mL insulin after three days. The medium was replaced every two days.
Adipogenic differentiation of HPA-s was carried out according to previous methods14 (link). HPA-s were cultured for 3 days followed by stimulating with differentiation medium for 5 days. The differentiation medium consists of 500 ml of PADM basal medium (Cat. No. 7221, ScienCell), 25 ml of fetal bovine serum (FBS, Cat. No. 0025, ScienCell), 5 ml of preadipocyte differentiation supplement (PAdDS, Cat. No. 7232, ScienCell), 5 ml of penicillin/streptomycin solution (P/S, Cat. No. 0503, ScienCell). The medium was replaced every two days.
3T3-L1 preadipocytes were cultured for 2 days, then 3T3-L1 cells were stimulated with differentiation medium for 3 days. The differentiation medium consists of DMEM (Cat. No.11965092, ThermoFisher) with 10% FBS and MDI (0.5 mM 1-methyl-3-isobutylxanthine (Cat. No. I5879, Sigma-Aldrich), 1 μM dexamethasone (Cat. No.D4902, Sigma-Aldrich), and 10 μg/mL insulin, Cat. No. 12585014, Sigma-Aldrich). Cells were cultured with DMEM containing 10% FBS and 10 μg/mL insulin after three days. The medium was replaced every two days.
5
Human Trabecular Meshwork Cell Culture
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All experiments were performed in an in vitro cellular model using commercially available HTM cells (6590) isolated from the juxtacanalicular and corneoscleral regions of the human eye. The cell line was purchased from the ScienCell Research Laboratories (San Diego, CA, USA). Cell culture was maintained under standard conditions (37 °C; 5% CO2; 95% humidity), according to the guidelines provided by the vendors. Cells were cultured in trabecular meshwork cell medium (TMCM) (ScienCell Research Laboratories, San Diego, CA, USA), that contains basic medium (BM) (ScienCell Research Laboratories, San Diego, CA, USA), 2% fetal bovine serum (FBS) (ScienCell Research Laboratories, San Diego, CA, USA), 1% trabecular meshwork cell growth supplement (TMCGS) (ScienCell Research Laboratories, San Diego, CA, USA) and 1% penicillin/streptomycin solution (P/S) (ScienCell Research Laboratories, San Diego ad, CA, USA). Cells were cultured on poly-L-lysine-coated T-75 culture vessels (2 μg/cm2). Cells were split every 3–4 days, when the culture reached 90–95% confluency, after exposure to 0.05% trypsin/EDTA solution (T/E) solution (ScienCell Research Laboratories, San Diego, CA, USA).
6
In Vitro Analysis of Dental Cements
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All of the in vitro analyses were performed in an experimental model using a commercially available monocyte/macrophage peripheral blood cell line—SC (ATCC CRL-9855) (ATCC; Manassas, VA, USA). Cell cultures were kept under standard conditions (37 °C; 5% pCO2; 95% humidity) according to the guidelines provided by the manufacturer. Cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) with 4-mML-glutamine adjusted to contain 1.5 g/L sodium bicarbonate (ATCC; Manassas, VA, USA) and supplemented with 0.05-mM 2-mercaptoethanol (Sigma-Aldrich Corp., St. Louis, MO, USA), 0.1-mM hypoxanthine and 0.016-mM thymidine (90%) (ATCC; Manassas, VA, USA), fetal bovine serum (10%) (ATCC; Manassas, VA, USA) and 1% penicillin/streptomycin solution (P/S) (ScienCell Research Laboratories, San Diego ad, CA, USA). Each cell culture was split when it reached 90–95% confluency. All of the tested cements were mixed according to manufacturer’s instructions and then cured in sterile hemi-sphere molds, r = 3.75 mm (surface area 1.33 cm2, volume of 140 μL). Immediately after reaching the setting time as provided by the manufacturer, the specimens were placed in Eppendorf tubes containing 1 mL of cell culture medium and were incubated for 24 and 48 h at 37 °C. The eluates were centrifugated for 5 min (2000 rpm) and then used for further analysis [33 (link)].
7
Culturing Human Astrocytes and Glioma Cells
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Human astrocytes (HA) and human glioma cell lines (U87, LN229, U251, A172 and B19) were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA). Human THP-1 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All cell line authentication service was applied and mycoplasma was also routinely tested. Astrocyte medium (AM) (Cat. #1801, ScienCell, USA) was used to culture human astrocytes, supplemented with 10 mL of fetal bovine serum (FBS) (Cat. #0010, ScienCell, USA), 5 mL of penicillin/streptomycin solution (P/S) (Cat. #0503, ScienCell, USA) and 5 mL of Astrocyte Growth Supplement (AGS) (Cat. #1852, ScienCell, USA). The glioma cell lines were cultured in DMEM Medium (Dulbecco’s Modified Eagle Medium, Gibco, USA) and THP-1 cells were cultured in RPMI 1640 medium with 10% FBS (FBS, BI serum, Israel) and cultured in a 37°C constant temperature incubator containing 5% CO2.
8
Culturing HUVECs and HAECs for Endothelial Studies
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HUVECs were purchased from Lifeline cell technology (C-12200, cat. no. 10171-906). HAECs were a kind gift from Dr. Lusis (UCLA, David Geffen School of Medicine). HUVECs and HAECs were cultured in Petri dishes or flasks coated with 0.2% gelatin type A (cat. no. 901771; MP Biomedicals, Santa Ana, CA, USA), in Endothelial Cell Medium (ECM, Cat.no. 1001, ScienCell, Carlsbard, CA. USA) containing 465 mL of basal medium, 25 mL of fetal bovine serum (FBS, Cat. no. 0025, ScienCell, Carlsbard, CA, USA), 5 mL of Endothelial Cell Growth Supplement (ECGS, Cat. no. 1052, ScienCell, Carlsbard, CA, USA) and 5 mL of penicillin/streptomycin solution (P/S, Cat. no. 0503, ScienCell, Carlsbard, CA, USA). Only HUVECs with less than 6 passages and HAECs with <15 passages were used in this study.
9
Cultivation of Human Cardiac Myocytes
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Human cardiac myocytes (HCMs) were cultured with cardiac myocyte medium (CMM) in poly-L-lysine-coated dishes and supplemented with 5% fetal bovine serum (FBS), 1% cardiac myocyte growth supplement (CMGS), and 1% penicillin/streptomycin solution (P/S) (all were purchased from ScienCell Research Laboratories, Inc., Carlsbad, CA, USA). Cells were maintained in an incubator with 5% CO2 atmosphere at 37 °C.
10
Isolation and Culture of HUVECs and HAECs
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Human umbilical vein endothelial cells (HUVECs) were obtained from collagenase-digested umbilical cord veins (65 (link)) and collected in M200 medium supplemented with LSGS (Cascade Biologics, Inc., Portland, OR) and 2% FBS (Atlanta Biologicals, Inc., Lawrenceville, GA). Human aortic endothelial cells (HAECs) were a kind gift from Dr. Lusis (UCLA, David Geffen School of Medicine). HUVECs and HAECs were cultured in Petri dishes or flasks coated with 0.2% gelatin type A (cat. no. 901771; MP Biomedicals, Santa Ana, CA, USA), in Endothelial Cell Medium (ECM, Cat.no. 1001, ScienCell, Carlsbard, CA. USA) containing 465 mL of basal medium, 25 mL of fetal bovine serum (FBS, Cat. no. 0025, ScienCell, Carlsbard, CA, USA), 5 mL of Endothelial Cell Growth Supplement (ECGS, Cat. no. 1052, ScienCell, Carlsbard, CA, USA) and 5 mL of penicillin/streptomycin solution (P/S, Cat. no. 0503, ScienCell, Carlsbard, CA, USA). Only HAECs with <15 passages were used in this study.
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