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Catalogue Number
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UBP-0031
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Product Format
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Frozen Vial
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Cell Number
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> 5×10[5]
cells/vial
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GENERAL INFORMATIONHBAs (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).
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CHARACTERIZATION OF
THE CELLS
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1.
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Cytoplasmic GFAP
> 98% positive by immunofluorescence
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2.
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HBAs are negative
for HIV-1, HBV, HCV, and mycoplasma
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PRODUCT USAGE Cells are offered for Research Use Only.
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SHIPPING Frozen Vials in a Dry Ice Package.
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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.
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PROTOCOLS FOR
THAWING THE CELLS AND SUBCULTURE
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A)
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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.
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B)
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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.
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C)
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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.
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D)
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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.
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E)
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Add 5ml Universal Neutralization Buffer and spin down the
cells with 800g centrifugation for 5 mins.
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G)
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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).
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H)
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Change medium every
2 or 3days and the cells usually become confluent within 7 days (when split
at a 1/3 ratio).
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I)
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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.
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