Male & female neonatal C57BL/6J pups (Jackson Laboratories, Bar Harbor, ME) were used for primary OPC cultures and female adult mice were used for the various in vivo demyelinating disease models (cuprizone, LPC, and C‐EAE). Male and female postnatal day 5 (P5) Sprague Dawley rat pups (Charles River) were used for astrocyte cell cultures. All procedures were performed with the approval of the Northwestern University, George Washington University, and New York University Institutional Animal Care and Use Committees and adhered to the NIH Guide for the Care and Use of Laboratory Animals.
C57bl 6j pups
Manufactured by Jackson ImmunoResearch
Sourced in Montenegro
C57BL/6J pups are a commonly used mouse strain in biomedical research. They are the most widely used inbred mouse strain and serve as a standard control in many experiments.
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Lab products found in correlation
6 protocols using c57bl 6j pups
1
Multi-species Cell Culture Models
2
Organotypic Slice Culture for AAV Infection
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Brains were dissected from decapitated postnatal p2-p4 C57BL/6 J pups (Cat: 000664, The Jackson Laboratories) and glued (Glue Loctite) to the chuck of a water-cooled vibratome (Leica VT1000A) and trimmed with a commercial shave razor. Under aseptic conditions, 300 μm-thick coronal sections were sliced and collected in sterile medium. The organotypic slices were then placed in a 0.4-μm membrane insert (Millipore PICM03050) in a 6-well plate and cultured in presence of Basal Medium containing HBSS 1X, 15% Horse serum, GlutaMax, 45% Glucose, Pen-strep, N2 supplement, and fresh PDGF (10 ng/ml) and maintained at 35 °C and 5% CO2. Slices were infected with the respective AAV after 3 days of culturing. After AAV infection, slices were cultured for 9–12 days, during which time media was replenished with half media changes every three days. Slices were collected and lysed with Lysis Buffer as described previously.
3
Isolation and Culture of O4+ OPCs
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O4+ OPCs were isolated by immunopanning and grown in culture as described previously (Rodgers et al., 2015 (link)). Male and female C57BL/6J pups (Jackson Labs) were used for experiments involving only wild type cells and in separate experiments, pups from a cross between Cnp+/+; Hcn2F/F and Cnp+/cre; Hcn2F/F animals were used. All pups were aged P7-9 for isolations.
4
Primary OPC and Astrocyte Cultures
5
Primary Muller Glia and RGC Cultures
6
Organotypic Slice Culture and AAV Infection
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Brains were dissected from decapitated postnatal p2-p4 C57BL/6J pups (Cat: 000664, The Jackson Laboratories) and glued (Glue Loctite) to the chuck of a water-cooled vibratome (Leica VT1000A) and trimmed with a commercial shave razor. Under aseptic conditions, 300μm-thick coronal sections were sliced and collected in sterile medium. The organotypic slices were then placed in a 0.4-μm membrane insert (Millipore, PICM03050) in a 6-well plate and cultured in presence of Basal Medium containing HBSS 1X, 15% Horse serum, GlutaMax, 45% Glucose, Pen-strep, N2 supplement, and fresh PDGF (10ng/ml) and maintained at 35°C and 5% CO2. Slices were infected with the respective AAV after 3 days of culturing. After AAV infection, slices were cultured for 9–12 days, during which time half of the media volume was replaced with one half fresh media every three days. Slices were collected and lysed with Lysis Buffer as described below.
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