All reagents were purchased from Thermo Fisher Scientific, MA, U.S.A. unless stated otherwise. Human primary fetal astrocytes here, referred to as FPA (ScienCell, U.S.A), were maintained as per the supplier’s instructions. In brief, the cells were cultured in tissue culture-treated T75 flasks coated with 1× Attachment Factor protein in ScienCell astrocyte media (AM) with the supplements of 2% FBS, 1% astrocyte growth supplement (AGS), and 1% PenStrep (all ScienCell). When confluent, the cells were passaged at a ratio of 1:4, using TrypLE incubation for 4 min. Defined Trypsin Inhibitor (DTI) was used for TrypLE inhibition; the cells were spun down to a pellet at 200 g, after which they were resuspended in the supplemented AM. The U87 glioblastoma cells, previously published by Pontén and MacIntyre [21 ] and obtained from Uppsala University, were maintained in tissue culture-treated T75 flasks in high glucose DMEM supplemented with 10% FBS and 1% PenStrep. When confluent, the cells were passaged at a ratio of 1:4 by a 4 min TrypLE incubation. The cells were then spun down to a pellet at 200 g, after which they were resuspended in the supplemented DMEM described above.
Pen strep
Manufactured by ScienCell
Sourced in United States
Pen/Strep is a solution containing penicillin and streptomycin, which are antibiotics commonly used to prevent bacterial contamination in cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Astrocyte growth supplement, by ScienCell (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
Human cortical astrocytes, by ScienCell (1 mentions)
Tryplexpress, by Thermo Fisher Scientific (1 mentions)
Trypsin ethylenediaminetetraacetic acid edta, by Thermo Fisher Scientific (1 mentions)
Dmem high glucose hyclone medium, by GE Healthcare (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Dulbecco s phosphate buffered saline dpbs, by Lonza (1 mentions)
5 protocols using pen strep
1
Culturing Primary Fetal Astrocytes and U87 Glioblastoma
2
Culturing Human Cortical Astrocytes
3
Culturing Mouse LSEC Cell Line
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4
Culturing HPV-immortalized Human Corneal Cells
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HPV-immortalized human corneal epithelial cells, a generous gift from Dr. May Griffith (Integrative Regenerative Medicine (IGEN) Centre, Linköping University, Sweden) [38] (link) were cultured in keratinocyte serum free medium (KSFM) supplemented with bovine pituitary extract, recombinant epidermal growth factor, and penicillin/streptomycin (Pen/Strep) (ScienCell, Carlsbad, California, USA) at and 5% carbon dioxide ( ). Fresh medium was added every other day and cells were grown to 90% confluency in tissue culture treated flasks. Adherent cells were removed using TryplExpress (Life Technologies, Burlington, Ontario, Canada) dissociation solution. Cells were routinely observed for any morphological changes and were used before their eleventh passage.
5
Culturing Pancreatic Stellate and Cancer Cells
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Pancreatic stellate cells (PSCs, ScienCell, Carlsbad, CA, USA) were cultured in stellate cell medium supplemented with 2 v/v % fetal bovine serum (FBS), 100 U/mL penicillin/100 µg/mL streptomycin (Pen/Strep) and 1 v/v % stellate cell growth supplement (SteCGS) according to manufacturer’s instruction (all products from ScienCell). Panc-1 cancer cells (ATCC, Manassas, VA, USA) were cultured in DMEM - High Glucose HyClone medium (GE Healthcare Life Sciences, Chicago, IL, USA) containing 10 v/v % FBS, 100 U/mL penicillin/100 µg/mL streptomycin (Thermofisher Scientific) and 2 mM L-glutamine (Thermofisher Scientific). The cells were maintained at 37 °C in a humidified 5% CO2 atmosphere and passed at 80% confluence. Passing of cells was performed as follows: The cells were washed twice with warm Dulbecco’s phosphate buffered saline (DPBS) (Lonza, Basel, Switzerland) before addition of trypsin/ethylenediaminetetraacetic acid (EDTA) (Thermofisher Scientific) and incubation at 37 °C. The trypsin/EDTA mix was neutralized by using 10× cell culture medium before being transported to a sterile Falcon tube and counted using a hemocytometer (Buerker-Tuerk, Brand GMBH, Wertheim, Germany). PSCs were used from a passage of 4–10.
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