Human Brain Microvascular Endothelial Cells Expressing GFP-AEQ in Mitochondria

Name of Product
Human Brain Microvascular Endothelial Cells Expressing GFP-AEQ in Mitochondria

Catalog Number 
HB02

Product Format
Frozen Vial

Cell Number

5 × 10⁵ cells/vial

Product Description
These cells were isolated from normal human brain cortex tissue and genetically modified to express GFP-AEQUORIN localized in the mitochondria. Selection was performed using puromycin after lentiviral infection. Cells are supplied at passage 1 and shipped in cryopreserved vials. Nuvyra Labs Endothelial Growth Medium is recommended for optimal culture conditions.

Storage & Stability

• Storage Temperature: –196°C (Liquid Nitrogen Vapor Phase)

• Shipping Temperature: –80°C

• Stability: Long-term stability maintained in the vapor phase of liquid nitrogen

Cell Characterization / Specification

• CVWF/Factor VIII (Cytoplasmic): >95% positive (Immunofluorescence)

• Cytoplasmic Di-I-Ac-LDL Uptake: >95% positive (Immunofluorescence)

• Cytoplasmic PECAM1 (CD31): >95% positive (Immunofluorescence)

Quality Control

Handling Upon Arrival
Transfer vials to a –80°C freezer for short-term storage or liquid nitrogen for long-term preservation.

Thawing & Subculturing Protocol

1. Pre-Coat Flask:
   Add 2 mL of Nuvyra Labs Coating Solution to a T25 flask. Incubate for 5 min. Aspirate, rinse with 4 mL cell rinse solution, and discard.

2. Thawing:
   Thaw vial in a 37°C water bath. Spray with 70% ethanol, wipe, and transfer to biosafety cabinet. Add 10 mL of Nuvyra Labs Endothelial Growth Medium to pre-coated flask and seed the cells.

3. Passaging:

   • Rinse flask with 5 mL rinse solution ×2.

   • Add 2 mL Detachment Solution. Aspirate after 20 seconds.

   • Incubate for 1–2 min at RT or 37°C. Tap gently to detach.

   • Add 5 mL Neutralization Solution and centrifuge at 800g for 5 min.

4. Seeding:
   Resuspend pellet in 10–15 mL medium. Distribute 5 mL per pre-coated T25 flask (1:2 or 1:3 split).
   Replace medium every 2–3 days. Confluence reached in ~7 days.

5. For Quiescent Cells:
   Once nearly confluent, switch to Endothelial Basal Medium for 12 hours before experiments.

Product Usage
Cells are offered for Research Use Only. Not for Clinical Use.