The murine macrophage cell line J774A.1 mCherry was purchased from ATCC and modified by René Neuhaus from Pallasch Lab (Cologne, Germany) using a mCherry MLS empty vector. The human-MYC/BCL2 (hMB) GFP+ cell line (strain 102) was generated by Leskov et al. [17 (link)]. Cells were cultured in Dulbeccos modified Eagels Medium (DMEM) (Cat.No. P04-03550, PAN-Biotech) supplemented with 10% Fetal Bovine Serum (Sigma) and 1% Penicillin-Streptavidin (Biochrom) in a 5% CO2 atmosphere with 95% humidity at 37 °C. Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages for in vitro experiments were isolated from C57BL/6JRcc mice. For differentiation into BMDMs, bone marrow-derived cells were put on 10 cm dishes with DMEM treated with 10 ng/ml recombinant murine M-CSF (Peprotech) for 7 days. Peritoneal macrophages were isolated via peritoneal lavage by injecting the peritoneum with 5 ml cold PBS, shaking the mouse for 2 min, then aspirating the injected fluid. Peritoneal lavage was centrifuged 5 min at 400 g, supernatant was discarded and the cell pellet was resuspended in DMEM for culturing. After culturing them for 2 h at 37 °C, the non-adherent cells were removed and adherend cells were supplemented with new media.
Penicillin streptavidin
Manufactured by Harvard Bioscience
Sourced in Germany
Penicillin-Streptavidin is a laboratory reagent used in various biochemical and molecular biology applications. It consists of the antibiotic penicillin covalently linked to the protein streptavidin. The core function of this product is to provide a means of specifically binding and detecting biotinylated molecules in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant murine m csf, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Insulin, by PromoCell (1 mentions)
Fetal calf serum (fcs), by Harvard Bioscience (1 mentions)
Dulbecco s modified eagle s medium, by PAN Biotech (1 mentions)
Epidermal growth factor (egf), by Roche (1 mentions)
Neonatal normal human epidermal keratinocytes, by Lonza (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Fetal calf serum (fcs), by PAN Biotech (1 mentions)
2 protocols using penicillin streptavidin
1
Murine and Human Cell Culture Protocols
2
Culturing Human Skin Cell Lines
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Neonatal Normal Human Epidermal Keratinocytes (Lonza, Walkersville, MD, U.S.) were cultured in Basal Medium 2 (PromoCell, Heidelberg, Germany) supplemented with Keratinocyte Growth Medium 2 Supplement Pack (PromoCell).
The A431 cell line (human cutaneous epidermoid carcinoma, female origin, RRID:CVCL_0037) was obtained from Cell Lines Services (Eppelheim, Germany) and cultured in Dulbecco's Modified Eagle's Medium with 4.5 g/l Glucose and L-Glutamine supplemented with 10% fetal calf serum (PAN-Biotech, Aidenbach, Germany).
The human cSCC cell line SCC-12 was kindly provided by Prof. Jens Malte Baron from Aachen, Germany (male origin, RRID:CVCL_4026). Cells were cultured in Ham´s F-2 Medium (Biochrom, Berlin, Germany) and Dulbecco's Modified Eagle's Medium (PAN-Biotech) in a 1:4-ratio, supplemented with 10% fetal calf serum, 1% Penicillin/Streptavidin (Biochrom), 40 ng/ml Hydrocortisone (Sigma-Aldrich, Darmstadt, Germany), 10 ng/ml EGF (Roche, Basel, Schweiz) and 5 µg/ml Insulin (PromoCell).
Cells were incubated at 37 °C in a humidified 5% carbon dioxide atmosphere.
The cell lines used differ in sex of donors. Since we did not analyze sex-specific differences, this can be viewed as a limitation concerning our analyses.
The A431 cell line (human cutaneous epidermoid carcinoma, female origin, RRID:CVCL_0037) was obtained from Cell Lines Services (Eppelheim, Germany) and cultured in Dulbecco's Modified Eagle's Medium with 4.5 g/l Glucose and L-Glutamine supplemented with 10% fetal calf serum (PAN-Biotech, Aidenbach, Germany).
The human cSCC cell line SCC-12 was kindly provided by Prof. Jens Malte Baron from Aachen, Germany (male origin, RRID:CVCL_4026). Cells were cultured in Ham´s F-2 Medium (Biochrom, Berlin, Germany) and Dulbecco's Modified Eagle's Medium (PAN-Biotech) in a 1:4-ratio, supplemented with 10% fetal calf serum, 1% Penicillin/Streptavidin (Biochrom), 40 ng/ml Hydrocortisone (Sigma-Aldrich, Darmstadt, Germany), 10 ng/ml EGF (Roche, Basel, Schweiz) and 5 µg/ml Insulin (PromoCell).
Cells were incubated at 37 °C in a humidified 5% carbon dioxide atmosphere.
The cell lines used differ in sex of donors. Since we did not analyze sex-specific differences, this can be viewed as a limitation concerning our analyses.
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