STRO-1, HOP-26 (CD63), CD49a and SB-10 (CD166) as markers of primitive human marrow stromal cells and their more differentiated progeny: a comparative investigation in vitro

Abstract

There is widespread interest in the use of bone marrow stromal cells (BMSC) for tissue reconstruction and repair and for gene therapy. BMSC represent the differentiated progeny of CFU-F, which however comprise a developmentally heterogeneous population as is reflected in the cellular heterogeneity of the cell populations to which they give rise. We have compared the efficacy of monoclonal antibodies recognising a series of stromal antigens, viz. STRO-1, HOP-26, CD49a and SB-10/CD166, as tools for the enrichment of CFU-F prior to culture and as developmental markers for culture-expanded BMSC. In freshly isolated bone marrow mononuclear cells (BMMNC), the proportion of antigen-positive cells was 27%, 46%, 5% and 19% for STRO-1, HOP-26, CD49a and CD166, respectively. All CD49a(+) cells co-expressed STRO-1. The degree of CFU-F enrichment obtained with anti-CD49a (approximately 18-fold) by a one-pass immunoselection strategy was significantly greater than that of all other antibodies tested. BMSC expressed higher levels of all antigens investigated (except for HOP-26) compared with BMMNC. Expression of STRO-1 and CD49a remained restricted to a subset of BMSC, whereas all BMSC were SB-10/CD166 positive. Treatment with dexamethasone (10 nM), which promotes the differentiation and further maturation of cells of the osteogenic lineage in this cell culture system, increased the expression of CD49a and HOP-26. The CD49a(+) and HOP-26(+) fractions of BMSC were further subdivided by dual-labelling with anti-STRO-1 and B4-78 (an antibody recognising the B/L/K isoform of the enzyme alkaline phosphatase), respectively. By using a variety of criteria, the HOP-26 antigen was identified as CD63, a member of the tetraspanin family of proteins thought to modulate integrin compartmentalisation and signalling.

Morphological, molecular and functional differences of adult bone marrow- and adipose-derived stem cells isolated from rats of different ages

“…Stro1 was used as a marker of undifferentiated stem cells in preference to other markers as it recognises nonhaematopoietic progenitor cell with a stable undifferentiated phenotype [35]. Also, we previously found that expression of other markers is inconsistent.…”

Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis

“…Especially, we found that CD18 is highly expressed on the BMSSCs of the early passages after their purification from bone marrow. Given such a distinct pattern of expression, CD18 could be a useful surface marker for cell sorting to enrich the stromal stem cell population, because most of the other surface markers for stromal stem cells, including STRO-1, CD106͞VCAM-1, CD146͞MUC-18, HOP-26, CD49A͞integrin ␤ 1 , and SB-10͞ CD166 (20,22,28), are expressed both on progenitor and on differentiated mesenchymal cells. As proof of principle, we isolated BMSSCs from human bone marrow aspirates by cell sorting using both CD18 and STRO-1, a cell marker commonly used for stromal stem cells (12).…”

Tissue engineering: strategies, stem cells and scaffolds

“…As marrow is a complex mixture of cells a more defined starting cell population subset can be isolated from the mixture on the basis of epitope expression such as Endoglin (Haynesworth et al 1992;Majumdar et al 2000) and STRO-1 (Simmons & Torok-Storb, 1991;Howard et al 2002;Stewart et al 2003) antibody selection procedures. These cells can be removed from marrow and used to enhance materials such as the filler used for stabilizing artificial hip joints or for joining critical sized defects in bone that would not otherwise heal (Tilley et al 2006).…”