Human blood and marrow side population stem cell and Stro-1 positive bone marrow stromal cell numbers decline with age, with an increase in quality of surviving stem cells: correlation with cytokines.

Hematological deficiencies increase with aging leading to anemias, reduced hematopoietic stress responses and myelodysplasias. This study tested the hypothesis that side population hematopoietic stem cells (SP-HSC) would decrease with aging, correlating with IGF-1 and IL-6 levels and increases in bone marrow fat. Marrow was obtained from the femoral head and trochanteric region of the femur at surgery for total hip replacement (N=100). Whole trabecular marrow samples were ground in a sterile mortar and pestle and cellularity and fat content determined. Marrow and blood mononuclear cells were stained with Hoechst dye and the SP-HSC profiles acquired. Marrow stromal cells (MSC) were enumerated flow cytometrically employing the Stro-1 antibody, and clonally in the colony forming unit fibroblast (CFU-F) assay. Plasma levels of IGF-1 (ng/ml) and IL-6 (pg/ml) were measured by ELISA. SP-HSC in blood and bone marrow decreased with age but the quality of the surviving stem cells increased. MSC decreased non-significantly. IGF-1 levels (mean=30.7, SEM=2) decreased and IL-6 levels (mean=4.4, SEM=1) increased with age as did marrow fat (mean=1.2mmfat/g, SEM=0.04). There were no significant correlations between cytokine levels or fat and SP-HSC numbers. Stem cells appear to be progressively lost with aging and only the highest quality stem cells survive.

Cardiolipin liposomes: a novel flow reagent for detection of anticardiolipin antibodies.

The association of autoantibodies with specificity for phospholipids and an increased risk for thromboembolic phenomena has received considerable recent clinical attention. These autoantibodies have been reported in patients with defined autoimmune disorders as well as in patients with no other obvious autoimmune disease symptoms other than isolated or recurrent thromboembolic disease. A significant component of this autoimmune response appears to be related to cardiolipin-directed antibodies. Most studies reported to date have used either an enzyme immunoassay or a radioimmunoassay for detection and quantitation of antiphospholipid antibodies. We have developed a novel flow cytometric assay for detection of anticardiolipin antibodies. The assay, by analogy to polystyrene microsphere assay, utilizes cardiolipin liposomes as solid-phase microspheres for antigen presentation. In comparison to enzyme-linked immunosorbent assay, the flow assay shows similar sensitivity by serum titration, has immunoglobulin class specificity, and is semiquantitative as currently designed. The flow assay is relatively easy to perform and should allow detection of other antiphospholipid specificities with tailoring of the phospholipid makeup of the liposomes.