Angiogenesis Investigation

Creative Biolabs is one of the leading-edge institutions and service providers in the field of therapeutic antibody development. Currently, we proudly present a unique technology platform to evaluate antibody potencies towards angiogenesis based on a range of cell-based assays.

Introduction to Angiogenesis

Angiogenesis is a multi-step, complicated process, which involves multiple cell types acting in concert to form new blood vessels. Various growth factors, enzymes, and extracellular matrix molecules can participate in this mechanism, particularly vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP-1, MMP-2, MMP-9), angiopoietin-1 and 2 (Ang-1, Ang-2). Major steps during angiogenesis can be characterized as follows:

• Cell proliferation
• Migration
• Differentiation
• Anastomosis and refinement

It constitutes a key part in many critical bioactivities in vivo, such as tissue repair, wound healing, and embryonic development. However, under pathologic conditions, angiogenesis can also contribute to disease progression, for instance, establishing a favorable microenvironment for tumor development, dissemination, and metastasis. Therefore, anti- and pro-angiogenesis therapies that intend to regulate aberrant angiogenic activities have proven the great potential for treating solid tumors, arteriosclerosis, diabetes, myocardial infarction, ulcer, limb ischemia, tissue ischemia, etc., many of which have already advanced to clinical applications.

Fig.1 Effect of angiogenesis stimulators and an inhibitor on tube formation. (Arnaoutova, 2010)

In an effort to accelerating the development of antibody drugs with angiogenesis mechanisms, Creative Biolabs provides two ready-to-use in vitro models for phenotypically mimicking in vivo angiogenesis:

Primary Cell Model

Co-culture of human umbilical vein endothelial cells (HUVEC) and normal human dermal fibroblasts (NHDF). When exposed to appropriate inducing agents, HUVEC can undergo a similar process that closely resembles in vivo angiogenesis (including initial proliferation) in 8-12 days.

Stem Cell Model

Co-culture of endothelial colony forming cells (ECFC) and adipose-derived stromal cells (ADSC). ECFC can recapitulate the major phases of angiogenesis except for initial proliferation, within 3-5 days. Notably, ADSC in this model can differentiate into pericytes (demonstrated by biomarkers of smooth muscle actin and PDGFR-β), which can support and nourish neonatal angiogenic networks.

Creative Biolabs has established a novel platform to real-time visualize and quantify phenotype changes in 96-well plate format. We also apply well-validated comparable controls (e.g. VEGF, Suramin, Sunitinib) to ascertain assay validity.

• Tube Formation: Our exclusive platform can real-time image and measure tube formation or disruption, like tube length, tube area, branch point formation over all phases of the angiogenesis process. Full kinetic data, as well as automated images, can be acquired.
• Biomarkers: We can investigate a full menu of relevant biomarkers (i.e. PECAM) using immunofluorescence staining, to further validate the presence of microvascular or certain cell types. Other specific markers are also available upon request.
• Cell Migration: During angiogenesis, cell migration plays a crucial role. To check whether an antibody can interfere with cell migration/invasion abilities is thereby of great importance. We use an automated scratch wound healing assay to obtain dynamic information about migration tendencies of target cells.
• In Vitro Cell Adhesion Assay: In vitro cell adhesion assay is an important, efficient experiment type to detect the response of tumor cells adhering to stromal cells or extracellular matrix, which is of great significance for discovering new drugs for treating cancer pathology. Creative Biolabs specializes in providing in vitro cell adhesion assay services with the potential to aid in cancer research.

Moreover, our highly professional researchers at Creative Biolabs are specialized in designing custom assays to satisfy every specific demand and expectations. We will work closely with you step by step to ensure the most reliable and relevant outcomes. For more detailed information, please feel free to contact us or directly sent us an inquiry.

Reference

1. Arnaoutova, I.; Kleinman, H. K. In vitro angiogenesis: endothelial cell tube formation on gelled basement membrane extract. Nature protocols. 2010, 5(4): 628.

For Research Use Only.