DESCRIPTION

BACKGROUND

Human ENPP1 (Ectonucleotide Pyrophosphatase/Phosphodiesterase 1) is a membrane-bound glycoprotein that belongs to the family of ectonucleotide pyrophosphatase/phosphodiesterase (ENPP) enzymes. ENPP1 is involved in a variety of physiological processes including bone mineralization, insulin signaling, and inflammation and is expressed in many tissues including bone, liver, adipose tissue, and cartilage. ENPP1 has both pyrophosphatase and phosphodiesterase activities and it plays a crucial role in regulating the levels of extracellular pyrophosphate, a mineralization inhibitor, in tissues such as bone and cartilage. It also serves as an inhibitor of insulin signaling, leading to insulin resistance in adipose tissue and skeletal muscle. In addition, ENPP1 has been implicated in various cellular processes such as cell adhesion, inflammation, and matrix calcification. ENPP1 has been found to be up regulated in various types of cancer, including breast, prostate, and colon cancer. Increased ENPP1 expression has been associated with cancer cell proliferation, migration, and invasion. This suggests that ENPP1 may play a role in cancer progression and metastasis. Given its link to a number of human diseases and cancers, there has been considerable interest in developing drugs that can modulate ENPP1 activity as a potential therapeutic strategy.

THAWING AND CULTURING

• Remove the cell vial from liquid nitrogen tank and thaw cells quickly in a 37oC water bath.

• In a T250 shake flask, add 30ml cell culture media which has been pre-warmed in a 37oC water bath for 30 minutes.

• Transfer all the cells in the vial into the T250 shake flask and place it on a shaker inside of a CO2 incubator with 5-8% CO2. The shaker speed should be set at 120-150rpm.

• The next day, count the cells. The viable cell density should be around 0.5-1x106/ml and the cell viability should be >50%.

• The cell doubling time should be 18-24 hours. Split the cells 1:5 or 1:10 every 2-3 days or when the cell density reached 6-8x106/ml.

DATA

Detection of human ENPP1 expression on human ENPP1-CHO-S stable cells using a monoclonal antibody specific for human ENPP1 (Accurus #A1022)

REFERENCES

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Roberts F, Zhu D, Farquharson C, Macrae VE. ENPP1 in the Regulation of Mineralization and Beyond. Trends Biochem Sci. 2019 Jul;44(7):616-628. doi: 10.1016/j.tibs.2019.01.010. Epub 2019 Feb 21. PMID: 30799235.

Bacci S, De Cosmo S, Prudente S, Trischitta V. ENPP1 gene, insulin resistance and related clinical outcomes. Curr Opin Clin Nutr Metab Care. 2007 Jul;10(4):403-9. doi: 10.1097/MCO.0b013e3281e386c9. PMID: 17563456.

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