Detection of the Hematopoietic Stem and Progenitor Cell Marker CD133 during Angiogenesis in Three-Dimensional Collagen Gel Culture

Abstract

We detected the hematopoietic stem and progenitor cell marker CD133 using immunogold labeling during angiogenesis in a three-dimensional collagen gel culture. CD133-positive cells were present in capillary tubes newly formed from aortic explants in vitro. The CD133-positive cell population had the capacity to form capillary tubes. Lovastatin strongly inhibited cell migration from aortic explants and caused the degradation of the capillary tubes. The present study provides insight into the function of CD133 during angiogenesis as well as an explanation for the antiangiogenic effect of statins.

Development of In Vitro Method for Assaying Anti-Angiogenic Effect of Drugs

“…Alexa Fluor 488-and Nanogold (1.4 nm)-conjugated goat anti-rabbit IgG (Nanoprobes, USA) were used as a secondary antibody. We proposed that CD133-positive cells related to the capillary tube elongation and/or branching [11]. In the study [11], CD133-positive cells were observed in the capillary tube (Fig.…”

“…CD133-positive cells were also presented within the aortic explant [11]. Human fetal aorta contained vascular progenitor cells.…”

“…Imaging was performed by a phase-contrast microscope (Nikon TE2000, Japan) with a stage preheated to 37 °C and an imaging system (Aquacosmos, Hamamatsu Photonics, Japan) [11]. The culture dish was maintained under 5% CO 2 during image acquisition.…”

“…We detected CD133 positive cells during angiogenesis in the collagen gel culture [11]. The samples were fixed in 4% paraformaldehyde/0.1 M phosphate buffer (pH 7.2) and treated with CD133 antibody (rabbit polyclonal; Abcam ® , Japan).…”

“…Endothelial precursor and stem cells are present in a zone of the vascular wall, which is located between the smooth muscle and the adventitial layer. It is suggested that the primary source of the capillary tube is the intimal endothelial cells [11]. These progenitor cells are present as a cluster between the outer smooth muscle layer and inner adventitia [118].…”