A detailed study of time‐dependent changes in human red blood cells: from reticulocyte maturation to erythrocyte senescence

Abstract

SummaryThe use of microfabrication technology in the study of biological systems continues to grow rapidly in both prevalence and ascendancy. Customised microdevices that provide superior results than traditional macroscopic methods can be designed in order to investigate specific cell types and cellular processes. This study showed the benefit of this approach in precisely characterising the progressive losses of surface area and haemoglobin (Hb) content by the human red blood cell (RBC), from newborn reticulocyte to senescent erythrocyte. The high-throughput, multiparametric measurements made on individual cells with a specialised microdevice enabled, for the first time, delineation and quantification of the losses that occur during the two stages of the human RBC lifespan. Data acquired on tens of thousands of red cells showed that nearly as much membrane area is lost during the 1-2 d of reticulocyte maturation (c. 10-14%) as in the subsequent 4 months of erythrocyte ageing (c. 16-17%). The total decrease in Hb over the red cell lifespan is also estimated (c. 15%) and a model describing the complete timecourse of diminishing mean RBC area and Hb is proposed. The relationship between the losses of Hb and area, and their possible influence on red cell lifespan, are discussed.

Maturing reticulocytes internalize plasma membrane in glycophorin A–containing vesicles that fuse with autophagosomes before exocytosis

“…10 Mature reticulocytes released from BM undergo final remodeling after release, resulting in a 10%-14% membrane loss in the first 48 hours. 29 Consistent with this, the circumference of cultured reticulocytes lacking staining for either LAMP1 or CD63 (30.92 Ϯ 2.31 m, n ϭ 8) was 10% less than those staining positive for either marker (33.96 Ϯ 2.23 m, n ϭ 9).…”

“…10 Mature reticulocytes released from BM undergo final remodeling after release, resulting in a 10%-14% membrane loss in the first 48 hours. 29 Consistent with this, the circumference of cultured reticulocytes lacking staining for either LAMP1 or CD63 (30.92 Ϯ 2.31 m, n ϭ 8) was 10% less than those staining positive for either marker (33.96 Ϯ 2.23 m, n ϭ 9).To determine the source of GPA in GPA-positive cytosolic vesicles, R2 reticulocytes were incubated at 37°C in the presence of anti-GPA, and the appearance of GPA-positive vesicles observed over time ( Figure 6A). The results clearly showed that GPA derives from the plasma membrane.…”

Extrinsic and intrinsic control by EKLF (KLF1) within a specialized erythroid niche

“…Erythroblastic islands were first observed in bone marrow, although they are also present in the splenic red pulp and in the fetal liver (Chasis and Mohandas, 2008;Manwani and Bieker, 2008). These islands consist of a central macrophage that extends cytoplasmic protrusions to a ring of 5-30 surrounding erythroid cells encompassing proerythroblast to reticulocyte stages (Gifford et al, 2006). Extensive macrophage-erythroblast and erythroblast-erythroblast adhesive interactions are necessary for a thriving definitive erythropoietic community.…”

Physiological and pathological population dynamics of circulating human red blood cells

“…Recently, it has become possible to identify and characterize very young (hours or days old) circulating RBCs (reticulocytes) (2). RBCs undergo a rapid reduction in volume and hemoglobin in the few days after release from the bone marrow (3). This rapid phase is followed by a much longer period of slower reduction (4-7) during which volume and hemoglobin are coregulated (8) (Fig.…”