hCMEC/D3

Name: hCMEC/D3
Brand: Procell
SKU: CL-0843
Price: 500.0 USD
Availability: InStock

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Cat. No.: CL-0843

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CL-0843

$ 500.00

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1×10^6Cells/Vial×2Vials

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Product Introduction Documents Publications

Product Introduction

General information

Cell Name	hCMEC/D3
Synonyms	HCMEC/D3;CMEC/D3;human Cortical Microvessels Endothelial Cells/D3
Cellosaurus Accession	CVCL_U985
Organism	Human
Growth Properties	Adherent
Morphology	Spindle-shaped
Tissue	Brain, temporal lobe, blood microvessel
Age	Adult
Sex	Female

Handling information

Instructions	1. Check all containers for leakage or breakage. 2. Remove the frozen cells from the dry ice packaging and immediately transfer them to liquid nitrogen (liquid or vapor phase) for long-term cryopreservation.
Complete Medium	Complete culture medium for hCMEC/D3
Incubation Atmosphere	Air, 95%; CO₂, 5%
Temperature	37℃
Subcultivation Ratio	1:3-1:6
Medium Renewal	2 to 3 times per week
Dissociation Duration	2-3 min
Subculturing Procedure	1. Remove the culture medium from the T25 cell culture flask. 2. Add approximately 2 mL of PBS. Gently tilt the flask side to side until the PBS covers the entire bottom, then aspirate and discard the PBS. 3. Add 1 mL of 0.25% trypsin solution (containing EDTA). Gently tilt the flask side to side until the trypsin solution covers the entire bottom of the flask. 4. Incubate the cells at 37°C. Observe the cells under an inverted microscope and terminate digestion once the cells round up and detach. To avoid clumping, do not agitate the cells by tapping or shaking the flask during detachment. 5. Add 3 mL of complete culture medium to terminate digestion and disperse into a single cell suspension. 6. Collect the cell suspension and centrifuge at 1200 rpm (approximately 250 ×g) for 3 minutes. Carefully aspirate and discard the supernatant. 7. Add fresh complete culture medium, pipette gently several times to resuspend the cells, and seed them at the appropriate ratio into a new culture flask. Loosen the cap or use a vented cap for incubation.
Freeze Medium	General Freezing Medium [PB180436]
Storage Conditions	For long-term cryopreservation, cryovials should be stored in liquid nitrogen at −150°C to −196°C. Storage at −80°C is restricted to short-term interim use only.

Reference materials

Background

The hCMEC/D3 cell line was derived from human temporal lobe microvessels isolated from tissue excised during surgery for control of epilepsy. The primary isolate was enriched in cerebral endothelial cells (CECs). In the first passage, cells were sequentially immortalized by lentiviral vector transduction with the catalytic subunit of human telomerase (hTERT) and SV40 large T antigen, following which CEC were selectively isolated by limited dilution cloning, and clones were extensively characterized for brain endothelial phenotype. This brain microvascular endothelial cell line represents one such model of the human BBB that can be easily grown and is amenable to cellular and molecular studies on pathological and drug transport mechanisms with relevance to the central nervous system (CNS).

Biosafety Level

BSL-2