The canine pleomorphic mammary adenoma cell line ZMTH3 [51 (link)] was cultured routinely in RPMI 1640 supplemented with 10% fetal calf serum (FCS) and 1% penicillin/streptomycin (Biochrom AG, Berlin, Germany). Rat granulosa cells (GFSHR-17) were cultivated in DMEM (Dulbecco’s Modified Eagle Medium) supplemented with 5% FCS, 1% penicillin/streptomycin (Biochrom AG, Berlin, Germany). The canine prostate adenocarcinoma cell line CT1258 was derived from an extremely aggressive canine prostate carcinoma [52 (link)] and cultured in Medium 199 (Life Technologies GmbH, Darmstadt, Germany) supplemented with 10% FCS and 2% penicillin/streptomycin (Biochrom AG, Berlin, Germany). The human ES cell line hES3 and the human iPSC line hCBiPS2 [53 (link)] were cultured and expanded on irradiated mouse embryonic fibroblasts (MEF) in knockout DMEM supplemented with 20% knockout serum replacement, 1mM L-glutamine, 0.1mM β-mercaptoethanol, 1% nonessential amino acid stock (all from Life Technologies) and 10ng/ml bFGF (supplied by the Institute for Technical Chemistry, Leibniz University Hannover). One day before laser transfection cells were detached from the feeder layer by 0.2% collagenase IV (Life Technologies) followed by an incubation step with TrypLE (Life Technologies) for single-cell dissociation and plated onto Matrigel™ (BD Biosciences) coated dishes in MEF-conditioned medium.
Penicillin streptomycin
Manufactured by Harvard Bioscience
Sourced in Germany, United States, United Kingdom, Austria, Italy, France
Penicillin/streptomycin is a sterile, ready-to-use solution containing the antibiotics penicillin and streptomycin. It is commonly used as a supplement in cell culture media to prevent bacterial contamination.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Harvard Bioscience (92 mentions)
L glutamine, by Harvard Bioscience (64 mentions)
Fetal calf serum (fcs), by Harvard Bioscience (38 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (30 mentions)
Dulbecco modified eagle medium (dmem), by Harvard Bioscience (24 mentions)
Glutamax, by Thermo Fisher Scientific (23 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (23 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (22 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (18 mentions)
Rpmi 1640 medium, by Harvard Bioscience (15 mentions)
Hepes, by Harvard Bioscience (13 mentions)
Non essential amino acids (neaa), by Harvard Bioscience (13 mentions)
Trypsin edta, by Harvard Bioscience (12 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (12 mentions)
Fetal bovine serum (fbs), by Merck Group (12 mentions)
Sodium pyruvate, by Harvard Bioscience (11 mentions)
Trypsin, by Harvard Bioscience (10 mentions)
L glutamine, by Merck Group (10 mentions)
L glutamine, by Thermo Fisher Scientific (10 mentions)
Insulin, by Merck Group (9 mentions)
426 protocols using penicillin streptomycin
1
Cultivation of Canine and Human Cell Lines
2
Cell culture protocols for endothelial and hepatoma cells
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Human umbilical vein EA.hy926 cells (ATCC CRL-2922) were grown in M199 medium containing 10% fetal bovine serum (FBS; Biochrom, Berlin, Germany), 1% penicillin/streptomycin (100 U/mL, Biochrom), 0.2% glutamine (200 mM, Lonza, Basel, Switzerland), 0.2% heparin (12.5 mg/mL, Carl Roth, Karlsruhe, Germany), and 0.6% ascorbic acid (20 mM, Sigma-Aldrich, Steinheim, Germany). HUVEC were freshly isolated from human umbilical cords and grown in M199 containing 17.5% FBS, 1% penicillin/streptomycin (100 U/mL), 0.34% glutamine (200 mM), 0.2% heparin (12.5 mg/mL), 0.5% ascorbic acid (20 mM) and endothelial mitogen (5 mg/mL, Alfa Aesar, Karlsruhe, Germany). FBS was heat inactivated at 56 °C. Rat hepatoma HTC4 cells expressing human S1PR1 together with human Gαi subunit of trimeric G proteins [15 (link)] were grown in minimal essential medium (MEM) with Earle’s salts (MEM Earle’s medium) containing 10% FBS, 2% 100× non-essential amino acids (Biochrom), 1% 100 mM sodium pyruvate (Biochrom) and 1% penicillin/streptomycin (100 U/mL). Cells were incubated at 37 °C and 5% CO2 in a humidified incubator (Panasonic, Hamburg, Germany).
3
Establishing Human Endothelial and Mesenchymal Stem Cell Cultures
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Human umbilical vein endothelial cells (HUVECs) (ATCC CRL-1730) were obtained from the American Type Culture Collection (Rockville, MD, USA) and cultured in low-glucose Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% fetal bovine serum (FBS, Cegrogen Biotech GmbH, Stadtallendorf, Germany) and 1% penicillin/streptomycin (Biochrom GmbH, Berlin, Germany). Cells were incubated at 37 °C in 5% CO2 in humidified air.
BMSCs were cultured in Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs (ATCC PCS-500-030), containing 10% FBS (Biochrom GmbH), 1% penicillin/streptomycin (Biochrom GmbH), and L-glutamine (4.5 mM) (Biochrom GmbH). Cells were incubated at 37 °C in 5% CO2 in humidified air.
BMSCs were cultured in Mesenchymal Stem Cell Basal Medium for Adipose, Umbilical and Bone Marrow-derived MSCs (ATCC PCS-500-030), containing 10% FBS (Biochrom GmbH), 1% penicillin/streptomycin (Biochrom GmbH), and L-glutamine (4.5 mM) (Biochrom GmbH). Cells were incubated at 37 °C in 5% CO2 in humidified air.
4
Cell-SELEX on Breast Cancer Cell Lines
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Human breast cancer MDA-MB-231 (ATCC HTB-26) and non-tumorigenic epithelial human breast MCF-10-2A (ATCC CRL-10781) cell lines were used for the cell-SELEX as target cells and negative control, respectively. Hs 578T (ATCC HTB-126), MDA-MB-468 (ATCC HTB-132), MDA-MB-453 (ATCC HTB-131), MCF-7 (ATCC HTB-22) and MDA-MB-157 (ATCC HTB-24) cell lines were also used to confirm the aptamers binding specificity. MDA-MB-231, Hs 578T, MDA-MB-468, MDA-MB-453, MCF-7 and MDA-MB-157 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) [Biochrom] supplemented with 10% Fetal Bovine Serum (FBS) [Biochrom] and 1% penicillin–streptomycin [Biochrom] on tissue culture treated flasks. MCF-10-2A cells were cultured in a 1:1 mixture of DMEM:Ham’s F12 medium [Biochrom] supplemented with 20 ng/ml Epidermal Growth Factor (EGF) [Sigma], 100 ng/ml cholera toxin [Sigma], 0.01 mg/ml insulin [Biochrom], 500 ng/ml hydrocortisone [Sigma], 5% horse serum [Biochrom] and 1% penicillin–streptomycin. Cells were propagated at 37 °C, 5% CO2 and 95% relative humidity. The cells were washed using phosphate-buffered saline (PBS) 1X pH 7.4 and were detached using Trypsin/EDTA solution [Biochrom]. Cells were counted in a hemocytometer using the trypan blue exclusion test.
5
Splenocyte Effect on Fibroblast Collagen
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To determine the impact of splenocytes on collagen production in fibroblasts, splenocytes of the different experimental groups were co-cultured with murine C4 fibroblasts. According to Van Linthout et al. (11 (link)), fibroblasts were plated at a density of 10,000 cells per well in Iscove Basal Medium (Sigma-Aldrich) containing 10% FBS and 1% penicillin/streptomycin (both Biochrom). Twenty-four hours later, isolated splenocytes were added at a ratio of 1 to 10 (fibroblasts to splenocytes) in Iscove medium (Sigma) supplemented with 10% FBS and 1% penicillin/streptomycin (both Biochrom) in the presence of 50 ng/ml of PMA and 500 ng/ml of Ionomycin (both BD Biosciences). After 24-h co-culture, splenocytes were removed, and fibroblasts were fixed with cold methanol. Subsequently, Sirius red staining was performed. For photometric analyses at 540 nm, the Spectra Max 340PC microplate reader (Molecular Device GmbH) was used.
6
Cell Culture Conditions for Induction of IDO and PD-L1
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A549 [25 (link)], BxPC3 [26 (link)], KATOIII [27 (link)], SKBR3 [28 ] cells were all grown in RPMI1640 medium (Biochrom AG) supplemented with 10% fetal calf serum (Biochrom AG), 1% penicillin/streptomycin (Biochrom AG), 50 μM 2-mercaptoethanol (Gibco), 1% nonessential amino acids (Biochrom AG), 1 mM sodium pyruvate (Biochrom AG) and 1 mM HEPES (Biochrom AG). SW480 [29 (link)] cells were grown in RPMI1640 medium supplemented with 10% fetal calf serum and 1% penicillin/streptomycin. A431 [25 (link)] cells were grown in DMEM (Biochrom AG) with 10% fetal calf serum and 1% penicillin/streptomycin. All cell lines were cultured according to standardized cell culture procedures. For induction of IDO and PD-L1 proteins, cells were treated for 48 h with 1000 U/ml IFNγ (Sigma-Aldrich).
7
Culturing Murine Cell Lines for L929 Conditioned Medium
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RAW264.7 cells (ATCC: TIB-71, murine macrophages) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Life Technologies, Darmstadt, Germany), supplemented with 10% FCS (PAA, Pasching, Austria) and 5% Penicillin/Streptomycin (Biochrom, Berlin, Germany). When cells reached a confluence of 70% they were scraped and split 1:4. NIH3T3 cells (ATCC: CRL-1658, murine embryonic fibroblasts) were grown in DMEM with GlutaMax™ (Life Technologies), supplemented with 10% FCS (PAA) and 5% Penicillin/Streptomycin (Biochrom). Every third day or at reaching a confluence of 50% cells were split 1:10. L929 cells (murine fibroblasts derived from C3H/An mice) were seeded at 5 x 105 on 75 cm2 cell culture flasks and cultured in 55 ml DMEM (high glucose, GlutaMAX™ Supplement, pyruvate) supplemented with 10% FCS and 5% Penicillin/Streptomycin for 10 days [38 (link)]. Medium was harvested, filtered through a 45μm filter system and transferred to 50 ml tubes. L929 conditioned medium was stored at -20°C.
8
Expansion of Transduced CD34+ Cells
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Transduced CD34+ cells were co-cultured with irradiated (80 Gy) murine fibroblasts that constitutively secrete human growth factors (M2-10B4, CLS Cell Lines Service). Before irradiation, M2-10B4 cells were grown in RPMI medium (Biochrom) with 10% FCS (FBS Superior, Biochrom) and 1% Penicillin-Streptomycin (Biochrom). CD34+ cells were cultured in Stem Span medium supplied with cytokines (human Flt3-Ligand, hFlt3 100 ng/ml, human stem cell Factor, hSCF 100 ng/ml, human thrombopoietin, hTPO 20 ng/ml, human interleukin 6, hIL-6 20 ng/ml, Peprotech) and 1% Penicillin-Streptomycin (Biochrom), through the transduction and sorting of transduced cells. Transduced CD34+ cells were grown for at least 6 weeks on irradiated M2-10B4 cells according to standard procedures in MyeloCult (Stem Cell Technologies). For optimal growth hydrocor-tisone (10−6 M, Stem Cell Technologies) and cytokines (hFlt3 20 ng/ml, hSCF 20 ng/ml, hTPO 4 ng/ml, hIL-6 4 ng/ml, Peprotech) were added. On a weekly basis, half the medium was changed and used for analysis.
9
Cell Culture Protocol for Tumor and Transduced Cell Lines
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Tumor cell lines, U-251 MG and EGFRvIII-expressing U-251 MG (U-251 MG/EGFRvIII), were described previously (16) . Both cell lines were cultured in DMEM (Gibco) containing 10% FBS (Invitrogen) and 100 U/mL penicillin/streptomycin (Gibco). Tumor cell lines, U-87 MG/EGFRvIII and GOS-3/EGFRvIII, were generated by transduction of U-87 MG tumor cells (ATCC) and GOS-3 [German Collection of Microorganisms and Cell Culture (DSMZ)] with human EGFRvIII cDNA, and were cultured in RPMI1640 (Biochrom) and DMEM containing 10% FBS, 100 U/mL penicillin/streptomycin, and 5 mg/mL Blasticidin (Gibco). The tumor cell line, DK-MG (DSMZ), was cultured in RPMI1640 containing 10% FBS, 100 U/mL penicillin/ streptomycin, and 1 Â nonessential amino acids (NEAA, Biochrom). CHO cells were transduced with wild-type EGFR (HER1), HER2, HER3, HER4, and human and cynomolgus monkey EGFRvIII. CHO cells were cultured in DMEM containing 10% FBS, 10 mmol/L sodium hypoxanthine, 1.6 mmol/L thymidine (1 Â HT Supplement, Gibco), and 1 Â NEAA (Biochrom). Transduced CHO cells were cultured in DMEM containing 10% FBS, 2 mmol/L L-Glutamine (Biochrom), 100 U/mL penicillin/streptomycin, and 1 Â NEAA. T-cell lines, HPB-ALL (DSMZ) and LnPx4119 (described previously; ref. 15) , were cultured in RPMI1640 containing 10% FBS and 100 U/mL penicillin/ streptomycin. All cells were cultured at 37 C in a 5% CO 2 chamber.
10
Establishment and Maintenance of Sarcoma Cell Lines
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Soft tissue sarcoma (STS) cell lines (SK-LMS, SW982, RD, SW872) as well as primary sarcoma cell lines (ssRMS, BR-CS and WW-LMS) isolated from consented rhabdomyosarcoma patients` primary tumors, were a gift of Dr. med. C. Hinterleitner and Prof. G. Kopp (University of Tübingen).
Cell lines SK-LMS, SW982, RD, ssRMS, BR-CS, and WW-LMS were maintained in Dulbecco’s Minimum Essential Media (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% penicillin–streptomycin (Biochrom), 1% Sodium pyruvate and 1% MEM-Non-Essential Amino acids (100X) (Gibco), while the SW872 was maintained in RPMI supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% penicillin–streptomycin (Biochrom), 1% Sodium pyruvate and 1% MEM-Non-Essential Amino acids (100X) (Gibco).
HEK239T cells used for lentiviral pseudo-virus production were obtained from ThermoFisher Scientific and maintained in Hyclone-DMEM medium supplemented with 10% FBS and 200 µM L-glutamine.
All cell lines were cultivated at 37 °C in 5% CO2 humidity.
Cell lines SK-LMS, SW982, RD, ssRMS, BR-CS, and WW-LMS were maintained in Dulbecco’s Minimum Essential Media (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% penicillin–streptomycin (Biochrom), 1% Sodium pyruvate and 1% MEM-Non-Essential Amino acids (100X) (Gibco), while the SW872 was maintained in RPMI supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% penicillin–streptomycin (Biochrom), 1% Sodium pyruvate and 1% MEM-Non-Essential Amino acids (100X) (Gibco).
HEK239T cells used for lentiviral pseudo-virus production were obtained from ThermoFisher Scientific and maintained in Hyclone-DMEM medium supplemented with 10% FBS and 200 µM L-glutamine.
All cell lines were cultivated at 37 °C in 5% CO2 humidity.
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