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Conditionally Immortalized Human Bone Marrow-Derived Mesenchymal Stem Cells

Size Price Quantity
1.0 Frozen Vial $4,500.00
Name of Products Conditionally Immortalized Human Bone Marrow-Derived Mesenchymal Stem Cells
Catalogue NumbercAP-0060
Product FormatFrozen Vial
Cell Number>5x10[5] cells/vial
HBMMSCs (cAP-0050) are isolated from human Bone Marrow tissues and demonstrated with spindle-shaped-, and fibroblast-like cells. CI-HBMMSCs (cAP-0060) are selected from Puromycin resistant HBMMSCs after infected with lentiviruses expressing SV40 LT under the control of TetON system. CI-HBMMSCs are maintained with proliferative capacity in the presence of Doxycycline (2ug/ml) in Mesenchymal Stem Cell Growth Medium (MSCGM, cAP-35).
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.
The cells are offered for Research Use Only.
Frozen Vials in a Dry Ice Package.
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.
A)Pre-coating of T25 flasks- Add 2ml each Quick Coating Solution (cAP-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 MSCGM (cAP-35) 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 Trypsin/EDTA (RT) (cAP-23) into one T25 flask; gently dispose the excessive Trypsin/EDTA 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 Trypsin 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).
1.To minimize the effect of SV40 LT antigen for your studies, cells could be cultured in the absence Doxycycline for 3-5 days before your final experiments.
2. The cells are growing better at 33C than 37C
3. The purchase of this product conveys to the buyer the nontransferable right to use the purchased item beyond buyâ¿¿s own Laboratory
Quick Coating SolutioncAP-01
MSC Growth MediumcAP-35
MSC Basal MediumcAP-35B
HBSS w/o Ca2+, Mg2+cAP-11
Cell Freezing Solution (FBS)cAP-22
Cell Freezing Solution (Non-FBS)cAP-22B
Trypsin/EDTA SolutioncAP-23
Trypsin Neutralization SolutioncAP-28
ITS (100x)cAP-26
L-Glutamine-MAXIMUM (100x) cAP-27
Human Plasma Fibronectin SolutioncAP-42
Handling human tissue derived products is potentially bio-hazardous, despite tested negative for HIV, HBV and HCV DNA; nonetheless, proper precautions must be taken to avoid inadvertent exposure.