Human Brain Microvascular Endothelial Cells

Catalog Number: UBP01BC
Product Format: Frozen Vial
Cell Number: >5 × 10⁵ cells per vial

BiologyCells’ Human Brain Microvascular Endothelial Cells are primary endothelial cells isolated from normal human brain microvasculature and supplied at passage one to preserve native microvascular endothelial phenotype, functional integrity, and experimental reliability. These cells provide a physiologically relevant in vitro model for blood–brain barrier research, neurovascular signaling, inflammation, angiogenesis, and central nervous system vascular studies.

Cells are supplied in frozen vial format to ensure stability, consistency, and reproducibility across experiments. Universal Endothelial Growth Medium is recommended for optimal attachment, growth performance, and maintenance of endothelial characteristics. The quality and growth features described are warranted only when BiologyCells-recommended media and reagents are used.

Storage and Shipping

Storage Temperature: −196 °C (liquid nitrogen vapor phase)
Shipping Temperature: −80 °C

Frozen vials are shipped on dry ice. Upon receipt, vials should be transferred immediately to a −80 °C freezer for short-term storage or to liquid nitrogen vapor phase storage for long-term preservation. Cells are very stable when stored in the liquid nitrogen vapor phase.

Safety and Handling

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

Due to the sensitive nature of primary cells and cell lines, all quality-related issues must be reported to BiologyCells within one month of product receipt. No quality warranty or replacement will be provided after this one-month period.

Cell Characterization and Quality Control

Each lot of Human Brain Microvascular Endothelial Cells undergoes rigorous quality control testing.

Endothelial Marker Expression (Immunofluorescence):
• Cytoplasmic von Willebrand factor / Factor VIII: >95% positive
• Cytoplasmic uptake of Di-I-Ac-LDL: >95% positive
• Cytoplasmic PECAM-1: >95% positive

Quality Testing Results:
• Sterility (bacteria): Negative
• Sterility (fungi): Negative
• Endotoxin level: <0.05 EU/µg (acceptance ≤1.0 EU/µg)
• Mycoplasma (PCR): Negative
• HIV-1 (PCR): Negative
• HIV-2 (PCR): Negative
• Hepatitis B (PCR): Negative
• Cell count: ≥5.0 × 10⁵ cells per vial
• Viability (Trypan Blue): >95% viable

Handling Upon Arrival

When the dry ice shipment is received, 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 coverage of the growth surface. Incubate at room temperature for five minutes. Aspirate excess coating solution, add cell rinse solution or one-times PBS, then remove the rinse. The flask is ready for use.

B) Thawing the cells
Thaw the frozen cell vial in a thirty-seven degree Celsius water bath. Spray the vial with seventy percent ethanol, wipe clean, and transfer 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 cultures become confluent, 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 to the flask and gently aspirate excess solution within twenty seconds.

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

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 seed 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

Human Brain Microvascular Endothelial Cells are intended for research use only and are not approved for diagnostic or therapeutic applications.