HPAEC-BMI1

HPAEC-BMI1 is a clonal cell line immortalized by stably expressing human BMI1 gene in primary pulmonary artery endothelial cells. 

The cells should be maintained in blasticidin (4 µg/mL) containing medium in routine cell culture.

Similar to hTERT immortalization techniques, BMI1-immortalized cells typically demonstrate the physiological characteristics of their parental primary cells. HPAEC-BMI1 cells retain important endothelial cell characteristics and functions such as express CD31/ PECAM-1, uptake acetylated low density lipoprotein (AcLDL), and form capillary-like tubes on a basement membrane matrix.

The base medium for this cell line is Vascular Cell Basal Medium (ATCC PCS-100-030). To make the complete growth medium, add the following components to the base medium:

• Endothelial Cell Growth Kit-BBE (ATCC PCS-100-040)
• 4 µg/mL blasticidin

To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.  

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 220 x g for 5 minutes.
4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO₂ in air atmosphere is recommended if using the medium described on this product sheet.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with D-PBS (ATCC 30-2200) to remove all traces of serum that contains trypsin inhibitor.
3. Add 3.0 to 4.0 mL of Trypsin-EDTA for Primary Cells solution (ATCC PCS-999-003) to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Once detached, add 3.0 to 4.0 mL of Trypsin Neutralizing Solution (ATCC PCS-999-004) and aspirate cells by gently pipetting. Transfer cell suspension into a 15mL conical tube.
5. Collect cells by centrifugation at 220 x g for 5 minutes. Discard the supernatant.
6. Resuspend cell pellet in 6.0 to 8.0 mL complete medium. Add appropriate aliquots of the cell suspension to new culture vessels.
   Cultures can be established between 4 x 10³ and 5 x 10³ viable cells/cm².
7. Incubate cultures at 37°C.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

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