Chromatin texture changes related to proliferation and maturation in erythrocytes

Abstract

Erythroid cell types were quantitatively assessed on the basis of the analysis of their nuclear images after Feulgen and Heidenhain blue counterstaining. An original nonsupervised classification method, developed in our laboratory, was applied to cell cycle analysis. Results obtained suggest that, in the four proliferative generation cycles, which concur with the successive morphological stages, there is a sequential flow of cells from one morphological stage to the next by cell cycling. We show that the chromatin pattern alone made it possible to set up a metric of the whole differentiation process occurring in the normal human erythroblastic lineage in which proliferating events occur. Key terms: Cell cycle analysis, differentiation in erythroblastic lineage, erythropoiesisErythropoiesis is an appropriate model for analyzing descriptors obtainable by means of high-resolution imcell cycle kinetic events and the maturation process in age analysis may become more accurate and convenient morphological changes during the evolution from prim-for functional interpretation (36,43,44). itive precursor cell to the mature erythrocyte, Differen-In a previous paper (261, we described a quantitative tiation of erythroid cells proceeds through a series of color image analysis of the normal maturation sequence recognizable morphological changes, which give rise to of the erythroblastic May-Grunwald-Giemsa (MGG) a cell type classification used by hematologists. The stained cells using the SAMBA 200 image analyzer. We subsequent stages of erythroid maturation are termed concluded that the machine successfully reproduced the mature erythrocyte is continuous although arbitrarily "he structural changes occurring during differentia-broken up by hematologists. tion are associated with striking biochemical alterationsThe present investigation deals with a combined quanas shown by combined methods of morphology and au-titative study of cell proliferation and cell differentiation toradiography (10,16,18,21,30,33,35), Unfortunately, of Feulgen Heidenhaim CFH1 blue-stained erythroid cells, these methods are time-consuming, and discrepancies in based on the concept that the functional properties of a the results reported were probably due to errors in the cell (DNA synthesis and template DNA activity) are experimental techniques used (34). A new approach to reflected in chromatin pattern. The nonsupervised this problem has been provided by analytical cytology method of classification (3,4) has the advantage of permethods based on computer-assisted cell-image process-mitting cell kinetic analysis from a single sample of a ing. The results already obtained by Zajicek et al. (46,471 cell population and does not require any a priori knowland Dormer et al. (17,19,20) for the characterization of edge about the proliferating state of the cells or a kinetic the grey-level images of erythroblasts at the various model of cell population. The results obtained will be steps of the maturation process have shown that it is discussed in terms of the coo...

Thalidomide alters nuclear architecture without ABCB1 gene modulation in drug-resistant myeloma cells

“…However, these variations in condensation are associated with modifications in chromatin spatial organisation. It has been proposed that chromatin texture could be described in terms of condensation (compactness of chromatin segments), distribution (relative proportions of compactness levels), and organisation (topographic arrangements of condensation degrees) (15). In 8226-Dox40 cells, significant variations were observed in E, and ENT, two features related to the spatial organization and distribution of the chromatin (15), and whose values are correlated neither to nuclear area nor to global chromatin condensation.…”

“…Four features were calculated on the gray-levels co-occurrence matrix : local mean of gray levels (LM), energy (E), entropy (ENT), and inertia (I). Five parameters were calculated on the run-length matrix: short run-length emphasis (SRE), long run-length emphasis (LRE), gray levels distribution (GLD), run-length distribution (RLD), and run-length percentage (RPC) (15).…”

From cytofluorometry to fluorescence image analysis.

“…Here, our novel approach has an important advantage that allows the morphometric analysis of the cells to be performed based on a biochemically specific fluorescence image, allowing the functional interpretation of cell morphology. In recent years, many studies using image analysis have been carried out to quantitatively analyze morphological and functional properties of the cells (6,7,11,14,18), but most of them are concerned with the cell images obtained with transmitted illumination.…”

Nuclear chromatin patterns in 3 glucocorticoid-resistant RPMI 8226 human myeloma cell sub-lines: Correlations with cell growth and immunological phenotype

“…Nine textural parameters were calculated on the nuclear image after reduction to 16 grey levels by linear rescaling. 19 Four features were calculated on the grey-levels cooccurrence matrix: Local Mean (LM), Energy (E), Entropy (ENT), and Inertia (I). Five parameters were calculated on the run-length matrix: Short Run-Length Emphasis (SRE), Long Run-Length Emphasis (LRE), Grey Levels Distribution (GLD), Run-Length Distribution (RLD), and Run-Length Percentage (RPC).…”

“…Five parameters were calculated on the run-length matrix: Short Run-Length Emphasis (SRE), Long Run-Length Emphasis (LRE), Grey Levels Distribution (GLD), Run-Length Distribution (RLD), and Run-Length Percentage (RPC). [19][20][21] The distribution, mean, and SD of the nuclear parameters were calculated for each cell population. Significance of the differences between parameters values was estimated by t test after Bonferroni correction for multiple variables.…”