The procedure for
isolating the astrocytes from neonatal rats was approved by the Institutional
Animal Care and Use Committee (IACUC) and completed at Wichita State
University in Wichita, Kansas.42 (link),43 (link) In brief, cerebral
cortexes were isolated from the brains of neonatal rats (postnatal
day P1-2 rats) after they were sacrificed. The cortex tissues were
triturated gently through a 5 mL syringe with a needle. The tissue
suspension was passed through a 70 mm nylon cell strainer (BD Falcon,
Durham, NC), and the flow-through was collected with a 50 mL conical
tube. The isolated cells were cultured for about 7–14 days.
After reaching confluency, the cultures were shaken to remove macrophages
and progenitor cells. The adherent astrocytes were cultured subsequently
for cell–nanofiber interaction. The cells were grown in Dulbecco’s
modified Eagle medium supplemented with 10% fetal bovine serum and
1% penicillin/streptomycin (Life Technologies, Grand Island, NY).
The medium was changed every 2–3 days, and the cell culture
was incubated with 5% CO2 at 37 °C.
isolating the astrocytes from neonatal rats was approved by the Institutional
Animal Care and Use Committee (IACUC) and completed at Wichita State
University in Wichita, Kansas.42 (link),43 (link) In brief, cerebral
cortexes were isolated from the brains of neonatal rats (postnatal
day P1-2 rats) after they were sacrificed. The cortex tissues were
triturated gently through a 5 mL syringe with a needle. The tissue
suspension was passed through a 70 mm nylon cell strainer (BD Falcon,
Durham, NC), and the flow-through was collected with a 50 mL conical
tube. The isolated cells were cultured for about 7–14 days.
After reaching confluency, the cultures were shaken to remove macrophages
and progenitor cells. The adherent astrocytes were cultured subsequently
for cell–nanofiber interaction. The cells were grown in Dulbecco’s
modified Eagle medium supplemented with 10% fetal bovine serum and
1% penicillin/streptomycin (Life Technologies, Grand Island, NY).
The medium was changed every 2–3 days, and the cell culture
was incubated with 5% CO2 at 37 °C.
