Perivascular cells expressing annexin A5 define a novel mesenchymal stem cell-like population with the capacity to differentiate into multiple mesenchymal lineages

Abstract

The annexin A5 gene (Anxa5) was recently found to be expressed in the developing and adult vascular system as well as the skeletal system. In this paper, the expression of an Anxa5-lacZ fusion gene was used to define the onset of expression in the vasculature and to characterize these Anxa5-lacZ-expressing vasculature-associated cells. After blastocyst implantation, Anxa5-lacZ-positive cells were first detected in extra-embryonic tissues and in angioblast progenitors forming the primary vascular plexus. Later, expression is highly restricted to perivascular cells in most blood vessels resembling pericytes or vascular smooth muscle cells. Viable Anxa5-lacZ+perivascular cells were isolated from embryos as well as adult brain meninges by specific staining with fluorescent X-gal substrates and cell-sorting. These purified lacZ+ cells specifically express known markers of pericytes, but also markers characteristic for stem cell populations. In vitro and in vivo differentiation experiments show that this cell pool expresses early markers of chondrogenesis, is capable of forming a calcified matrix and differentiates into adipocytes. Hence, Anxa5 expression in perivascular cells from mouse defines a novel population of cells with a distinct developmental potential.

Mice Lacking the Extracellular Matrix Protein WARP Develop Normally but Have Compromised Peripheral Nerve Structure and Function

“…Ultra thin sections were analyzed by transmission electron microscopy (LEO 906E, Oberkochen; Germany) as described previously (13).…”

Sorting of growth plate chondrocytes allows the isolation and characterization of cells of a defined differentiation status

“…Sections of Col10a1-LacZ transgenic mice were stained for b-galactosidase activity, as described previously. (23) Isolation of growth plate chondrocytes Distal femurs and proximal tibias of 12-or 13-day-old wild-type (C57BL/6), Col9a1-deficient or BAC-Col10a1-LacZ mice (15) were isolated and predigested with 0.2% collagenase P (Roche, Berlin, Germany) in DMEM and 10% fetal calf serum (FCS) for 30 minutes at 378C. Growth plate cartilage was prepared and digested with 0.2% collagenase II (Worthington) in DMEM and 10% FCS for 12 hours at 378C.…”

“…b-Galactosidase activity was detected by fluorescein-di-b-D-galactopyranoside staining as described previously. (23) Cells were subsequently washed and costained with 7AAD for dead-cell detection, followed by flow cytometry (FACSCalibur, FACSCantoII, Becton Dickinson) or cell sorting (FACSVantageSE, Becton Dickinson). For viability and cell-cycle experiments, cells were additionally stained with AnxA5-Alexa647 or Vybrant dye cycle green (Invitrogen) as described previously.…”

“…To isolate viable hypertrophic chondrocytes, growth plate cell suspensions were stained in parallel with the b-galactosidase substrate fluorescein (FIC)-di-b-D-galactopyranoside. (23) Using this analytical method, we identified a population of Col10a1-LacZ-FIC þ cells (9%, Fig. 2I) that also expresses the cell surface marker CD200 (Fig.…”

A Case of Extra-Skeletal Intracranial Chondromass: Ultrastructure and Immunohistochemical Features

“…It had been established that the MSC are important regulators of angiogenesis and blood vessel function [7] they also contribute to the pathogenesis of diabetic microangiopathy, cancer, atherosclerosis and Alzheimer's disease [6]. Furthermore the MSC pericytes have been reported to be able to differentiate into osteoblasts [8,9] chondrocytes and adipocytes [10,11].…”