Catalog Number: BCS11
Product Format: Frozen Vial
Cell Number: >5 × 10⁵ cells per vial

BiologyCells’ Human Aortic Artery Endothelial Cells are primary endothelial cells isolated from normal human aortic tissue and supplied at passage one to preserve native endothelial phenotype and functional integrity. These cells are intended for research applications involving cardiovascular biology, vascular function, endothelial signaling, and related in vitro studies.

Cells are provided in frozen vial format to ensure stability, consistency, and long-term usability. When cultured using BiologyCells-recommended endothelial growth media and reagents, the cells exhibit reliable attachment, characteristic endothelial morphology, and reproducible growth performance.

Storage and Shipping

Frozen vials are shipped on dry ice. Upon receipt, vials should be transferred immediately to a minus eighty degree Celsius freezer for short-term storage or to liquid nitrogen vapor phase storage at minus one hundred ninety-six degrees Celsius for long-term preservation. Cells remain highly stable when stored in the liquid nitrogen vapor phase.

Safety and Handling

Human tissue-derived products are potentially biohazardous, even when tested negative for HIV, HBV, and HCV DNA. Universal laboratory precautions must be followed when handling these materials. Appropriate personal protective equipment, including gloves and eye protection, should be worn at all times.

Due to the sensitive nature of primary cells, any quality-related concerns must be reported to BiologyCells within one month of product receipt. Quality claims submitted after this period are not eligible for replacement.

Cell Characterization and Quality Control

Each lot undergoes rigorous quality control testing. Cells demonstrate greater than ninety-five percent positivity by immunofluorescence for cytoplasmic von Willebrand factor and factor VIII, cytoplasmic uptake of Di-I-Ac-LDL, and cytoplasmic PECAM-1 expression.

Additional testing confirms:
• Negative sterility results for bacteria and fungi
• Endotoxin levels below 0.05 EU/µg
• Negative PCR results for mycoplasma, HIV-1, HIV-2, and hepatitis B
• Post-thaw viability exceeding ninety-five percent

Handling Upon Arrival

Upon receiving the dry ice shipment, frozen vials should be transferred immediately to a minus eighty degree Celsius freezer for short-term storage or to a liquid nitrogen tank for long-term storage.

Thawing and Subculturing Protocol

A) Pre-coating of T-25 flasks
Add two milliliters of Universal Coating Solution to a T-25 flask, ensuring full surface coverage. Incubate at room temperature for five minutes. Aspirate excess coating solution, rinse with cell rinse solution or one-times PBS, and remove the rinse. The flask is now ready for use.

B) Thawing the cells
Thaw the frozen vial in a thirty-seven degree Celsius water bath. Disinfect the vial with seventy percent ethanol and transfer it to a biosafety cabinet. Add ten milliliters of Universal Endothelial Growth Medium to the pre-coated T-25 flask and transfer the cells into the flask. Once cells reach confluency, they are ready for passaging.

C) Passaging the cells
Rinse cells twice with five milliliters of cell rinse solution. Add two milliliters of Universal Detachment Solution and aspirate excess solution within twenty seconds.

D) Detachment
Incubate the flask at room temperature or thirty-seven degrees Celsius for one minute. Cells typically detach within one to two minutes. Monitor microscopically and gently tap the flask to assist detachment if needed.

E) Centrifugation
Add five milliliters of Universal Neutralization Solution and centrifuge at eight hundred times gravity for five minutes.

F) Re-seeding
Re-suspend the cell pellet in ten to fifteen milliliters of growth medium and transfer five milliliters into two or three pre-coated T-25 flasks for a one-to-two or one-to-three split ratio.

G) Maintenance
Change medium every two to three days. Cells typically reach confluency within seven days when split at a one-to-three ratio.

H) Preparation for experiments
To prepare quiescent cells, replace growth medium with Universal Endothelial Basal Medium for approximately twelve hours once cultures are nearly confluent.

Product Use Notice

These Human Aortic Artery Endothelial Cells are for research use only and are not intended for diagnostic or therapeutic applications.