Human Retinal Pigment Epithelial Cells

Category:

Description

Name of Products Human Retinal Pigment Epithelial Cells
Catalogue Number UBP-0037
Product Format Frozen Vial
Cell Number > 5×10[5] cells/vial
GENERAL INFORMATIONHRPECs (UBP-0037) are initiated by digestion of minced human retinal pigment tissue with collagenase. HRPECs are separated from the mixture of cell populations and offered at passage 3 in a frozen vial. HRPECs Growth Medium (contains 10% serum and growth supplements, UBP-33) is recommended for cell culture and these cells have a minimum average population doubling Capacity > 8 when cultured following the detailed protocol described below).
CHARACTERIZATION OF THE CELLS
1. CRALBP Positive
2. RPE-65 Positive
3. HRPECs are negative for HIV-1, HBV, HCV, and mycoplasma.
PRODUCT USAGE: The 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 (although solution containing other extracellular matrix, i.e. gelatin, collagen, and fibronectin, can be used, make sure to optimize the conditions in advance).
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-33 medium, cells usually become confluent with 5-7 days.
C) To passage the cells, rinse the cells in a T25 flask with 5ml HBSS (RT) twice; then add 2ml Universal Coating Solution (RT) (UBP-23) into one T25 flask; gently dispose the excessive Universal Coating 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 Full 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).