Catalog Number RFP2
Product Name RFP Expressing Human Brain Microvascular Endothelial
Storage 37°C C0? incubator
Product Format Proliferating culture
Cells Number >90% confluent in T25 flask
*Caution: The handling of human derived products has the potential to be biologically hazardous. All Cell strains tested negative for HIV, HBV,
and HCV DNA in diagnostic tests. Proper precautions must be taken to avoid exposure. Always wear proper protective equipment (Gloves,
safety glasses, etc.) when handling these materials. We recommend following the universal procedures for handling products of human origin
as the minimum precaution against contamination.
RFP expressing human Brain Microvascular Endothelial Cells were isolated from normal human brain
tissue and transfected with RFP-Expressing lentiviral particles at passage 1. Puromycin resistant, RFPHBMVPCs is shipped in proliferating culture or frozen vial with a confluence of > 90% (cells are provide
@ passage 4-5). Endo-Growth medium (EGM2102) containing 5% serum and growth supplement is
recommended for culture. Cells have an average population doubling level >10 when cultured. When
you receive the cells, leave the flask in 37°C C02 incubator for 1 hour. Then, replace the transport
medium with fresh Endo-Growth medium (EGM2102). Let the cells grow for 24 hours before
Cytoplasmic Alpha-Actinin Filaments >80% positive by immunofluorescence
Cytoplasmic Desmin Intermediate Filaments >80% positive by immunofluorescence
Cytoplasmic Uptake of Di-I-Ac-LDL <2% Positive by immunofluorescence
Cytoplasmic VWF/ Factor VIII <2% positive by immunofluorescence
RFP expressing human brain microvascular
endothelial cells are negative for
HIV-1, HBV, HCV, and mycoplasma
PRODUCT USE AND SHIPPING STATUS
Product Use RFP expressing human brain microvascular
Endothelial cells are for research use only
Shipping Status Proliferating culture in T25 Flask
*Coating T25 flasks. Add 2 ml AlphaBioCoat (AC001) into 3- T25 flasks and ensure entire interior
surface is coated with the solution. After 30 minutes, dispose of AlphaBioCoat (AC001) by
aspiration. Gently rinse and aspirate the flask with Phosphate Buffer Solution (1XPBS-001). The
flask is now ready for use (no need for overnight incubation when coated with AC001). Add
fresh media to flask, if color changes from pink to yellow, discard the media, and add fresh
media to each flask.
1. Inspect to make sure Flask is at 90% confluence, if not remove transport media, and add 5ml of
fresh media to the flask. Place flask in 37°C incubator until cells are at 90% confluence. Change
media every 2 days.
2. If flask is at 90% confluence, aspirate transport media from flask.
3. Rinse T25 flask containing cells with 5 ml 1XPBS.
4. Gently aspirate out the PBS after rinsing, and discard.
5. Add 2ml of to T25 flask AlphabioDetach Cell Detachment Solution (ADF001) containing cells
(ensure entire interior surface is covered).
6. Place T25 flask containing cells into 37°C incubator for 1 or 2 minutes (cells will normally come
off of the surface within 1 or 2 minutes).
7. Suspend the cells with 15ml of Endo-Growth medium and transfer equally into 3 pre-coated T25
flasks (the cells are now at a subculture ratio of 1:3.
8. There is no need to spin cells during subculture.
9. Proliferating cells culture: Endo-Growth medium (EGM2102) should be changed every 2 days.
The cells normally become confluent within 7 days (when split at a 1:3 ratio).
10. Use Endo-Basal media (EBM-002) containing 0.5% FBS to induce quiescent cells (after 18-24