Long-term time-lapse cinemicrography of human erythroblasts under compression and without compression--direct measurement of generation time of each maturation stage.
The in vitro proliferation and the successive morphological alterations of single human erythroblasts were recorded for 5 days by time-lapse, phase-contrast cinemicrography. To visualize their fine structures more clearly, we introduced a new device to compress the erythroblasts under a cover slide. Appropriate compression did not significantly affect the capacity of the erythroblasts to proliferate and differentiate, although compression produced a slight tendency towards shorter generation time. The directly measured generation time was 24.5 +/- 4.5 h (mean +/- S.D.) for generation -1 (one generation prior to the orthochromatic erythroblast stage); 18.4 +/- 3.0 h for generation -2; and 18.3 +/- 4.1 h for generation -3 [corrected], these being arranged in inverse order of cell lineage. The cell size became progressively smaller and the nucleus-to-cytoplasm ratio decreased along with the maturation of the erythroblasts, showing a distinctive distribution range for each generation. Progression of hemoglobinization was simultaneously recorded qualitatively by Soret band absorption micrography. Cell movements characteristic for each maturation stage, e.g. perimitotic bleb-throwing of immature erythroblasts and vigorous movements of orthochromatic erythroblasts, heralding enucleation, were also clearly recorded. The method described here will be useful for the direct study of the dynamic morphology of the erythron in normal and abnormal states.
Inoue
H
H
Second Department of Pediatrics, Dokkyo University School of Medicine, Tochigi, Japan.
Sugita
K
K
Furukawa
T
T
D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D004900:Erythroblasts / Q000166:cytology*; D006801:Humans; D008853:Microscopy / Q000379:methods; D011312:Pressure; D013997:Time Factors