Reticulocyte Information and Courses from MediaLab, Inc.

Although the nucleus has been extruded, the reticulocyte is still considered immature because it retains numerous organelles needed for hemoglobin production, such as ribosomes, mitochondria, and fragments of the Golgi apparatus. The reticulocyte is slightly larger (10 microns) than the mature erythrocyte. A reticulocyte normally remains in the bone marrow for one or two days before entering the circulation and its final 24 hours of maturation. The red cell is mature when hemoglobin production is complete and the organelles have disintegrated. Reticulocytes normally make up 0.5 - 1.5% of the peripheral blood red cells. They appear blue/gray on the Wright's stained smear. The residual RNA in the cytoplasm causes the blue/gray color. The terms, polychromasia or polychromatophilic, are used to describe these cells on a Wright's stained preparation. A supravital stain such as new methylene blue N or brilliant cresyl blue is used to stain reticulocytes for an actual count.

When the large reticulocytes normally found in the bone marrow are present in the peripheral blood, they are referred to as shift or stress reticulocytes. These cells may be up to twice the size of normal mature red cells and are an indication of the bone marrow’s response to severe anemia. In addition to recognizing their appearance as polychromatophlic cells on Wright’s stained smears, it is now possible to quantify stress reticulocytes using a flourescent stain. They are classified as high, medium or low using a fluorescent-sensitive flow cytometer.

Erythrocyte production (including reticulocytes) is increased when the tissues are not receiving sufficient oxygen and the bone marrow is able to respond in a positive manner.Erythrocyte production (including reticulocytes) is decreased when the bone marrow is unable to respond to the signal for increasing production.

The large red cell in the center of this image is an example of a "shift" or stress reticulocyte. This cell has entered the circulation earlier than normal.

These dark blue or blue-purple particles are predominantly ribosomes (RNA), normally present in reticulocytes, which have aggregated. It is thought that basophilic stippling is not present in living cells but rather is produced during the smear preparation or staining process.

Erythrocyte inclusion is a term used to describe structures that may be present in red cells. The inclusions discussed in this course are: reticulocyte - Wright-Giemsa stain (polychromasia)basophilic stipplingHowell-Jolly bodiesPappenheimer bodies (Wright-Giemsa stain)siderotic granules (iron stain)Cabot rings

Jaundice was recognized in a day-old infant. Notice particularly the size variation (anisocytosis) of the erythrocytes on the infant's peripheral smear. What does this observation mean? Does it provide immediate information that might serve as guidance in expediting diagnosis and treatment? Note that normal-sized red blood cells, microcytes, microspherocytes, macrocytes, and nucleated red blood cells are all present. Red cell variations are expected findings in healthy neonates, but the variations here are exaggerated. Hyposplenic functional features may appear, including acanthocytes, spherocytes, and possibly Howell-Jolly bodies, especially if hemolysis is particularly vigorous. A high (3-7%) reticulocyte count is not unusual during the first three or four days after birth, however, the marrow in this jaundiced infant is proliferating vigorously in response to hemolysis. A call for more red cells is urgent. Immature red cells (in the form of nucleated red cells) and red cells with stippling of RNA (basophilic stippling) are readily identified. Red cell maturation sequence has not been totally processed in the marrow nor is all residual red cell debris removed by the spleen. In the lower photograph are reticulocytes stained by supravital stain (new methylene blue). Basophilic stippling (specks of RNA) stains with both supravital stains and with routine Wright-Giemsa stain.

Illustrated in the photomicrograph of a peripheral smear are two populations of erythrocytes. Approximately 50% of the erythrocytes are normal size and contain a full complement of hemoglobin. The patient had received blood transfusions. The transfused red blood cells are the normocytic, normochromic red cells. Admixed are microcytic erythrocytes and larger erythrocytes, some faintly mottled or smudged, suggestive of reticulocytes. This picture represents a hemolytic process with a reticulocyte response. A similar dimorphic red cell population appears following erythropoietin therapy. It is important to recognize when a population of cells in the peripheral smear is not in context with anticipated laboratory findings and the clinical situation.

In addition to the amount of hemoglobin present, the color of the cell must also be considered. Completely mature red cells appear buff-colored, while slightly immature non-nucleated red cells (reticulocyte stage) appear blue/gray on Wright's stained smears due to the presence of residual ribonucleic acid (RNA).The terms used to describe these cells are polychromasia or polychromatophilia. Polychromatophilic cells are frequently larger in size than mature red cells and can be distinguished from both types of macrocytes by this distinctive color.