Vascular cell adhesion molecule-1 expression and hematopoietic supportive capacity of immortalized murine stromal cell lines derived from fetal liver and adult bone marrow.
Ontogeny-specific differences in hematopoietic behavior may be influenced by unique adhesive interactions between hematopoietic cells and the microenvironment, such as that mediated by vascular cell adhesion molecule-1 (VCAM-1, CD 106). Although VCAM-1 is variably expressed during vertebrate development, we hypothesized that VCAM-1 expression might be linked to the enhanced capacity of the fetal liver microenvironment to support hematopoiesis. To test this we used immortalized murine stromal cell lines derived from midgestation fetal liver and adult bone marrow to compare the functional expression of VCAM-1. Molecular analysis of VCAM-1 expression was performed on stromal cell lines using Northern blot analysis, immunoprecipitation studies, and solid-phase enzyme-linked immunosorbent assay. Hematopoietic studies were performed by coculturing fetal liver cells with stromal cell lines, and the functional readout was determined by high-proliferative potential colony-forming cell (HPP-CFC) adherence assays. In contrast to our initial hypothesis, we observed greater expression of VCAM-1 messenger ribonucleic acid and protein on an adult marrow stromal cell line. In functional studies, anti-VCAM-1 antibody inhibited the binding of nearly half of the HPP-CFCs to adult marrow stroma but had a minimal effect on their binding to fetal liver stroma, despite the greater adherence of HPP-CFCs to fetal stroma. We conclude that VCAM-1 influences the hematopoietic supportive capacity of immortalized murine stroma derived from adult bone marrow. Our studies suggest that cellular interactions other than those mediated by VCAM-1 are involved in the increased adhesive capacity of immortalized murine stroma derived from fetal liver.
Koenig
Joyce M
JM
Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030, USA. koenijm@peds.ufl.edu
Ballantyne
Christie M
CM
Kumar
Ajith G
AG
Smith
C Wayne
CW
Yoder
Mervin C
MC
D000818:Animals; D000911:Antibodies, Monoclonal; D015152:Blotting, Northern; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D006224:Cricetinae; D005786:Gene Expression Regulation / Q000502:physiology*; D006410:Hematopoiesis; D008099:Liver / Q000166:cytology / Q000196:embryology; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017154:Stromal Cells / Q000502:physiology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis / Q000235:genetics*
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D019010:Vascular Cell Adhesion Molecule-1