1. New Information on Endothelial Cells
 
Pre-screened HUVEC, HMVEC and HAOEC VEGF
 
Signaling Pathway Tested
Endothelial cells are important parts of blood and lymph vessels. They play a pivotal role in a diverse range of physiological and pathophysiological processes from working as selective barrier between the blood and the surrounding bodily tissue to regulation of angiogenesis. They are closely involved in arterial disease and cancer development.



The most commonly used sources of human endothelial cells to study specific characteristics are HUVEC (Human Umbilical Vein Endothelial Cells) isolated from the vein of the umbilical cord, HMVEC (Human Microvascular Endothelial Cells) isolated from capillaries of the dermis and HAOEC (Human Aortic Endothelial Cells) isolated from capillaries of the aorta. At Cell Applications, primary cultures of HUVEC and HMVEC were pre-screened for angiogenesis and other endothelial cell signaling studies.


VEGF-Stimulated Signaling in Pre-Screened HUVEC, HMVEC and HAOEC
VEGF receptor-2 (VEGFR-2) is a major receptor transducing the signal of VEGF into the cells. Its expression is almost exclusively restricted to cells of endothelial lineage. VEGFR-2-mediated signaling plays a critical role in regulation of proliferation, differentiation, cell movement, and survival of endothelial cells and angiogenesis (1). VEGF-induced receptor dimerization triggers activation of VEGFR-2 tyrosine kinase and autophosphorylation at a specific set of tyrosine residues, which serve as docking sites for downstream signaling components containing SH2 or PTB domains. A regulated cascade of protein-protein interaction lead to activation of important signaling pathways within cells, such as Akt and MAP kinase activation (2, 3).

Detection of Expression of Important Endothelial Cell Signaling Molecules in Pre-Screened HUVEC and HMVEC
Tie2 is another important receptor tyrosine kinase critical for functional expression of endothelial cells and regulation of angiogenesis. Coordinated expression of Tie2 and its ligand angiopoietins is required for the angiogenic remodeling and vessel stabilization processes that occur subsequently to the initial vasculogenic action of VEGF receptors (4). It was demonstrated that Tie2-mediated signaling can effect changes in angiogenic sprout formation and survival of endothelial cells. Thus, Tie2 is also an important anti-angiogenesis drug target (5). The expression of Tie2 was detected in CAI pre-screened HUVEC and HMVEC cells as shown in Fig. 2, panel A.

Nitric oxide (NO) is an important signaling molecules that acts in many tissues to regulate a diverse range of physiological events and participates in various disease processes (6). NO is produced by the endothelial nitric oxide synthase (eNOS). Activation of eNOS leads to production of NO in endothelial cells, which regulates functional expression of endothelial cells in vascular remodeling and angiogenesis as well as cell survival (7). Abnormal control of NO synthesis has been implicated in a variety of diseases. Thus eNOS is an important endothelial cell signaling component target for drug development. The expression of eNOS was detected in CAI pre-screened HUVEC and HMVEC cells as shown in Fig. 2, panel B.

Axl is a new receptor tyrosine kinase family member. The survival factor growth arrest-specific gene 6 product (Gas6) is Axl ligand. Axl and Gas6 are both expressed in endothelial cells, where they are involved in many processes such as leukocyte transmigration through capillaries and neointima formation in injured vessels (8). Gas6 bound to HUVECs, protected HUVECs from apoptosis in response to growth factor withdrawal and from TNFalpha-mediated cytotoxicity. It was suggested Gas 6-Axl signaling may be relevant to endothelial cell survival in the quiescent environment of the vessel wall (9, 10). In addition, Axl stimulation by Gas6 results in inhibition of the ligand-dependent activation of vascular endothelial growth factor (VEGF) receptor 2 (11). There is a cross talk between Axl and VEGF receptor 2 in angiogenesis regulation. The expression of Axl receptor tyrosine kinase was detected in CAI pre-screened HUVEC and HMVEC cells as shown in Fig. 2, panel C.

The endothelial/epithelial tyrosine kinase (Etk/Bmx), a member of the Btk nonreceptor tyrosine kinase family, has been implicated in cell adhesion, migration, proliferation, and survival. Etk participates in signal transduction stimulated by growth factor receptors, cytokine receptors, G-protein-coupled receptors, antigen receptors, and integrins (12, 13). Etk has been implicated in the signaling of Tie-2 and VEGF receptors (14). Recently, it was demonstrated Etk is one of key players in regulation of TNF-induced inflammatory angiogenesis (15). The expression of Etk/Bmx tyrosine kinase was detected in CAI pre-screened HUVEC and HMVEC cells as shown in Fig. 2, panel D.

Product Name Product Description Size (vol) Cat. No.

Pre-Screened HUVEC Total Kit

neonatal

Each kit contains an ampoule of cryopreserved HUVEC (S200-05n), 500 ml of Endothelial Cell Growth Medium (211-500), and a Subculture Reagent Kit (090K). 1 Kit S200K-05n

Cryopreserved Pre-Screened HUVEC

neonatal

(>500,000 cells in an ampoule)

Cryopreserved in ampoules at the end of primary culture in Endothelial Cell Basal Medium that contains 10% FBS and 10% DMSO.

1 Ampoule S200-05n

Proliferating Pre-Screened HUVEC , neonatal 

In Flasks

Cultured in flasks at first passage in Endothelial Cell Growth Medium and shipped in Endothelial Cell Transfer Medium.

T-25 Flask

2 T-25 Flasks

T-75 Flask or 3 T-25 Flasks

S201-25n

S201-50n

S201-75n

Proliferating Pre-Screened HUVEC , neonatal  

In Multiwell Plates

Cultured in plates at first passage in Endothelial Cell Growth Medium and shipped in Endothelial Cell Transfer Medium.

6 well 

12 well

24 well

48 well

96 well

S201-6Wn

S201-12Wn

S201-24Wn

S201-48Wn

S201-96Wn

Endothelial Cell Growth Medium Optimized and ready for use 500 ml 211-500
Endothelial Cell Growth Medium Kit Basal Medium & Growth Supplement packaged separately Yields 500 ml 211K-500
Endothelial Cell Basal Medium Add Growth Supplement before use 500 ml 210-500
Endothelial Cell Growth Supplement   15 ml 211-GS
Subculture Reagent Kit 100ml each of HBSS, Trypsin/EDTA & Trypsin Neutralizing Solution 100 ml 090K

Pre-Screened CADMEC™ Total Kit

adult

neonatal

Each kit contains an ampoule of cryopreserved Pre-Screened CADMEC™ (S100-05), 500 ml of CADMEC™ Cell Growth Medium (112-500), a Subculture Reagent Kit (090K), two Extracellular Matrix Attachment Factor Coated T-25 Flasks (121-25-02), and 100 ml of Attachment Factor Solution (123-100). 1 Kit S100K-05a

S100K-05n

Cryopreserved Pre-Screened CADMEC™

adult

neonatal

(>500,000 cells in an ampoule)

Cryopreserved at second passage in CADMEC™ Cell Basal Medium that contains 10% FBS and 10% DMSO.

1 Ampoule S100-05a

S100-05n

Proliferating Pre-Screened CADMEC™

In Flasks

Cultured in flasks at third passage in CADMEC™ Growth Medium and shipped in Endothelial Cell Transfer Medium.

 

*Specify adult (a) or neonatal (n)

T-25 Flask

2 T-25 Flasks

T-75 Flask or 3 T-25 Flasks

S101-25*

S101-50

S101-75

Proliferating Pre-Screened CADMEC™

In Multiwell Plates

Cultured in plates at third passage in CADMEC™ Growth Medium and shipped in Endothelial Cell Transfer Medium.

 

*Specify adult (a) or neonatal (n)

6 well 

12 well

24 well

48 well

96 well

S101-6W*

S101-12W

S101-24W

S101-48W

S101-96W

CADMEC™ Growth Medium Optimized and ready for use 500 ml 112-500
CADMEC™ Growth Medium Kit Basal Medium & Growth Supplement packaged separately Yields 500 ml 112K-500
CADMEC™ Basal Medium Add Growth Supplement before use 500 ml 110-500
CADMEC™ Growth Supplement   30 ml 112-GS
Subculture Reagent Kit 100 ml each of HBSS, Trypsin/EDTA & Trypsin Neutralizing Solution 100 ml 090K