PARP-1 Knockout Immortalized Mouse Heart Endothelial (HYKO6) Cell Line
|T3031||1x106 cells / 1.0 ml|
|Description||Poly(ADP-ribose) polymerase 1 (PARP-1) is an ADP-ribosylating enzyme that plays a critical role in DNA damage repair, in addition to functioning as a context- specific regulator of transcription factors. Various pathophysiological conditions, such as reperfusion injury, endotoxic shock, diabetes and aging are associated with endothelial cell dysfunction mediated by PARP-1. |
The PARP-1-/-Knockout Immortalized Mouse Heart Endothelial (HYKO6) Cell Line is derived from stable transformation of SV40 into primary mouse heart endothelial cells from mice deficient for PARP-1. This cell line is capable of morphologic differentiation in capillary-like structures (CLS), similar to its primary counterparts, as shown by branching and anastomosing cords of cell networks and tube formation after 12 hours culturing on Matrigel. Moreover, HYKO6 responds to cytokine stimulation and has been shown to exhibit increased VCAM-1 and E-selectin expression after TNFα stimulation, making this cell line an important tool in the elucidation of PARP-1 signaling pathway involved in endothelial cell dysfunction and molecular mechanisms controlling cell gene expression in response to inflammatory stimuli.
|Species||Mouse (M. musculus)|
|Species description||Mouse (C57BL/6J)|
|Seeding Density||Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.|
For Research Use Only
|Unit quantity||1x106 cells / 1.0 ml|
|Caution||For Research Use Only|
|Cell Type||Drug Discovery Cells|
CD105, CD31, ICAM-2, VCAM-1, vWF
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37.0°C.
|Preservation Protocol||1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.|
2. Storage Temperature: Liquid nitrogen vapour phase.
1) Flow cytometry was used to confirm the expression of endothelial cell markers such as CD105, CD31, ICAM-2, VCAM-1 and MHC Class I antigens; 2) Immunofluorescence staining was used to assess the vWF expression; 3) Western blot was used to evaluate the expression of SV40 T-antigen in the immortalized cell line.
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3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
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- Carrillo, A et al. "Establishment of an immortalized PARP-1-/- murine endothelial cell line: a new tool to study PARP-1 mediated endothelial cell dysfunction" J Cell Biochem 94(6):1163-74 (2005). PubMed: 15696577.