(UBP-0031) are isolated from normal human brain cortical tissue. The cells are shipped in frozen vials (the cells are provided @ passage 1). Astrocytes Growth Medium (AGM, contains FBS and Growth factor supplements, UBP-29) is recommended for cell culture and these cells have a minimum average population doubling levels > 10 when cultured following the detailed protocol described below).
1.Cytoplasmic GFAP > 98% positive by immunofluorescence
2.HBAs are negative for HIV-1, HBV, HCV, and mycoplasmaPRODUCT USAGE Cells are offered for Research Use Only.SHIPPING Frozen Vials in a Dry Ice Package.
HANDLING OF ARRIVING CELLS When you receive the dry ice package with cells in frozen vials, transfer the frozen vials of cells into a -80C freezer for short period storage or a liquid nitrogen tank for long- term storage.PROTOCOLS FOR THAWING THE CELLS AND SUBCULTURE
A) Pre-coating of T25 flasks- Add 2ml each Universal Coating Solution (UBP-01) into a T25 flask to cover the whole surface of the flask, 5 mins later, dispose the excessive coating solution by aspiration and the flask is ready to be used.
B)Thaw the frozen cell vial in a 37C water bath first, and then transfer the cells into the pre-coated T25 flask with 10ml of UBP-44 medium, cells usually become confluent with 1-2 days and ready to be passaged.
C)To passage the cells, rinse the cells in a T25 flask with 5ml HBSS (RT) twice; then add 2ml Universal Detachment Solution (RT) (UBP-23) into one T25 flask; gently dispose the excessive Universal Detachment Solution within 20 seconds by aspiration.
D)Leave the T25 flask with the cells at RT or 37C for 1 min (most cells usually will detach from the surface within 1-2 mins; or monitor the cells under a microscope until most of cells become rounded up, and then gently tap the flask against the bench surface, and the cells will move on the surface of the flask when monitoring under microscope.
E)Add 5ml Universal Neutralization Buffer and spin down the cells with 800g centrifugation for 5 mins.
G)Re-suspend the cell pellet with 10 or 15ml UBP-09 medium and transfer 5 ml each into 2 or 3 pre-coated T25 flasks (for 1/2 to 1/3 subculture ratio).
H)Change medium every 2 or 3days and the cells usually become confluent within 7 days (when split at a 1/3 ratio).
I)To prepare quiescent cells, when cells are nearly confluent, replace UBP-29 with Pericyte Basal Medium (ABM, UBP-29B) containing 0.5%FBS for about 8-12hrs before your experiments.