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RFP Human Bone Marrow-Derived Mesenchymal Stem Cells

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Product Description
Name of Products RFP Human Bone Marrow-Derived Mesenchymal Stem Cells
Catalogue Number UBP-0050RFP
Product Format Frozen Vial
Cell Number > 5×10[5] cells/vial
GENERAL INFORMATION: RFP HBMMSCs (UBP-0050) are isolated from human Bone Marrow tissues and demonstrated with spindle-shaped-, and fibroblast-like cells. Each lot of UBP-0050 is tested to ensure the cells can be passaged at least three times (i.e., approximately 9 to 10 population doublings) after thaw in complete growth media (Mesenchymal Stem Cell Growth Medium (MSCGM), UBP-35), when cultured following the detailed protocol described below).
CHARACTERIZATION OF THE CELLS
1. Positive for CD29, CD44, CD73, CD90, CD105, and CD166 (greater than 95% of the cell population expresses these markers by flow cytometry).
2. Negative for CD14, CD31, CD34, and CD45 (less than 2% of cell population expresses these markers by flow cytometry).
3. HBMMSCs are negative for HIV-1, -2, HBV, HCV, and Bacteria, Yeast 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
HBMMSCs are contact inhibited. It is essential that the cells be subculture BEFORE reaching confluence as post-confluent cells exhibit changes in morphology, slower proliferation, and reduced differentiation after passaging.
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-35 medium, cells usually become confluent with 1-2 days.
C) To passage the cells, rinse the cells in a T25 flask with 5ml HBSS (RT) twice; then add 2ml Universal Detaching Solution (RT) (UBP-23) into one T25 flask; gently dispose the excessive Universal Detaching 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).
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