Rapidly Self-Renewing Human Multipotent Marrow Stromal Cells (hMSC) Express Sialyl Lewis X and Actively Adhere to Arterial Endothelium in a Chick Embryo Model System

Abstract

BackgroundThere have been conflicting observations regarding the receptors utilized by human multipotent mesenchymal bone marrow stromal cells (hMSC) to adhere to endothelial cells (EC). To address the discrepancies, we performed experiments with cells prepared with a standardized, low-density protocol preserving a sub-population of small cells that are rapidly self-renewing.MethodsSialyl Lewis X (SLeX) and α4 integrin expression were determined by flow cytometry. Fucosyltransferase expression was determined by quantitative realtime RT-PCR. Cell adhesion assays were carried out with a panel of endothelial cells from arteries, veins and the microvasculature in vitro. In vivo experiments were performed to determine single cell interactions in the chick embryo chorioallantoic membrane (CAM). The CAM is a well-characterized respiratory organ allowing for time-lapse image acquisition of large numbers of cells treated with blocking antibodies against adhesion molecules expressed on hMSC.ResultshMSC expressed α4 integrin, SLeX and fucosyltransferase 4 and adhered to human EC from arteries, veins and the microvasculature under static conditions in vitro. In vivo, hMSC rolled on and adhered to arterioles in the chick embryo CAM, whereas control melanoma cells embolized. Inhibition of α4 integrin and/or SLeX with blocking antibodies reduced rolling and adhesion in arterioles and increased embolism of hMSC.ConclusionsThe results demonstrated that rapidly self-renewing hMSC were retained in the CAM because they rolled on and adhered to respiratory arteriolar EC in an α4 integrin- and SLeX-dependent manner. It is therefore important to select cells based on their cell adhesion receptor profile as well as size depending on the intended target of the cell and the injection route.

Freezing Nitrogen Ethanol Composite May be a Viable Approach for Cryotherapy of Human Giant Cell Tumor of Bone

“…The reliability of the chicken chorioallantoic membrane model has been validated for investigation of the progression and invasiveness of human GCT [32]. It also has been affirmed that rapid self-renewing human multipotent mesenchymal bone marrow stromal cells actively adhere to arterial endothelium in a chicken chorioallantoic membrane model [3,21]. In particular, the chicken chorioallantoic membrane model has been used for evaluating the efficacy of pharmacologic treatment on GCTs by determining the level of angiogenesis [32].…”

Chorioallantoic Membrane Assay as Model for Angiogenesis in Tissue Engineering: Focus on Stem Cells

“…Several research groups showed that application of bone marrow-derived MSCs (BM-MSCs) on the CAM induced neovascularization. 89,[102][103][104] Oskowitz et al demonstrated that this effect was enhanced after serum deprivation of the cells during 30 days. 89 Similarly, MSCs isolated from adipose tissue (AT-MSCs), Wharton's Jelly (WJ-MSCs), placenta (Pl-MSCs), newborn skin (NSSCs), and soft tissues in and around the tooth penetrated the chicken embryo and increased functional blood vessel growth in the CAM 15,22,78,92,93,[105][106][107][108][109][110][111] (Table 1).…”

“…108,112 Furthermore, immune-based staining against human antigens is applied to determine the viability and potential incorporation and differentiation of the stem cells into newly formed blood vessels. 92,[110][111][112] Another regularly used method to track transplanted stem cells in the CAM is their prelabeling with fluorescent dyes such as CellTracker Green 104,107,111 or Fluoresbrite carboxylate microspheres. 103 Advanced imaging techniques.…”

Coalescent angiogenesis—evidence for a novel concept of vascular network maturation

“…The chick chorioallantoic membrane (CAM) presents a versatile in vivo model for the study of angiogenesis and vascular adaptation [ 30 ] as it facilitates optical access to a developing vascular system in a transparent, two-dimensional layout. Importantly, the CAM allows for extended observation periods for up to 1–2 days, permitting to follow the successive steps of angiogenic development over time [ 31 ]. In the fertilized chicken egg, the CAM is created through fusion of the allantois and the chorion from day 3 of embryonic development and its development continues until embryonic day 13 [ 32 ].…”