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Basic Characterization of the IMG-A1 Granulosa Cell Line. ( a , b ) Phase-contrast micrographs of the IMG-A1 cell culture. ( c ) Staining for senescence-associated β-galactosidase activity in passage 18 cells. ( d – f ) Immunocytochemical staining for the proliferation marker Ki67, showing a phase-contrast image ( d ), Ki67 immunofluorescence ( e ), and a merged image with DAPI nuclear counterstain ( f ). ( g , h ) Representative metaphase spreads showing a normal diploid karyotype (40 chromosomes) ( g ) and a near-tetraploid karyotype (78 chromosomes) from the IMG-A1 line ( h ). ( i – k ) Ploidy analysis by flow cytometry. Control blood cells ( i ) and primary granulosa cells ( j ) show characteristic diploid (2n) and tetraploid (4n, G2/M phase) peaks. The IMG-A1 culture ( k ) displays a predominantly near-tetraploid and near-octaploid cell population. ( l ) Flow cytometry analysis confirming the homogeneity of the IMG-A1 culture (passages 10 and 40) based on staining for the stromal marker CD29 and the absence of hematopoietic markers CD45, <t>CD34,</t> and the fibroblast marker CD90. Quadrant gates were set based on unstained controls. Scale bars = 100 µm.
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Basic Characterization of the IMG-A1 Granulosa Cell Line. ( a , b ) Phase-contrast micrographs of the IMG-A1 cell culture. ( c ) Staining for senescence-associated β-galactosidase activity in passage 18 cells. ( d – f ) Immunocytochemical staining for the proliferation marker Ki67, showing a phase-contrast image ( d ), Ki67 immunofluorescence ( e ), and a merged image with DAPI nuclear counterstain ( f ). ( g , h ) Representative metaphase spreads showing a normal diploid karyotype (40 chromosomes) ( g ) and a near-tetraploid karyotype (78 chromosomes) from the IMG-A1 line ( h ). ( i – k ) Ploidy analysis by flow cytometry. Control blood cells ( i ) and primary granulosa cells ( j ) show characteristic diploid (2n) and tetraploid (4n, G2/M phase) peaks. The IMG-A1 culture ( k ) displays a predominantly near-tetraploid and near-octaploid cell population. ( l ) Flow cytometry analysis confirming the homogeneity of the IMG-A1 culture (passages 10 and 40) based on staining for the stromal marker CD29 and the absence of hematopoietic markers CD45, <t>CD34,</t> and the fibroblast marker CD90. Quadrant gates were set based on unstained controls. Scale bars = 100 µm.
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Basic Characterization of the IMG-A1 Granulosa Cell Line. ( a , b ) Phase-contrast micrographs of the IMG-A1 cell culture. ( c ) Staining for senescence-associated β-galactosidase activity in passage 18 cells. ( d – f ) Immunocytochemical staining for the proliferation marker Ki67, showing a phase-contrast image ( d ), Ki67 immunofluorescence ( e ), and a merged image with DAPI nuclear counterstain ( f ). ( g , h ) Representative metaphase spreads showing a normal diploid karyotype (40 chromosomes) ( g ) and a near-tetraploid karyotype (78 chromosomes) from the IMG-A1 line ( h ). ( i – k ) Ploidy analysis by flow cytometry. Control blood cells ( i ) and primary granulosa cells ( j ) show characteristic diploid (2n) and tetraploid (4n, G2/M phase) peaks. The IMG-A1 culture ( k ) displays a predominantly near-tetraploid and near-octaploid cell population. ( l ) Flow cytometry analysis confirming the homogeneity of the IMG-A1 culture (passages 10 and 40) based on staining for the stromal marker CD29 and the absence of hematopoietic markers CD45, <t>CD34,</t> and the fibroblast marker CD90. Quadrant gates were set based on unstained controls. Scale bars = 100 µm.
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Basic Characterization of the IMG-A1 Granulosa Cell Line. ( a , b ) Phase-contrast micrographs of the IMG-A1 cell culture. ( c ) Staining for senescence-associated β-galactosidase activity in passage 18 cells. ( d – f ) Immunocytochemical staining for the proliferation marker Ki67, showing a phase-contrast image ( d ), Ki67 immunofluorescence ( e ), and a merged image with DAPI nuclear counterstain ( f ). ( g , h ) Representative metaphase spreads showing a normal diploid karyotype (40 chromosomes) ( g ) and a near-tetraploid karyotype (78 chromosomes) from the IMG-A1 line ( h ). ( i – k ) Ploidy analysis by flow cytometry. Control blood cells ( i ) and primary granulosa cells ( j ) show characteristic diploid (2n) and tetraploid (4n, G2/M phase) peaks. The IMG-A1 culture ( k ) displays a predominantly near-tetraploid and near-octaploid cell population. ( l ) Flow cytometry analysis confirming the homogeneity of the IMG-A1 culture (passages 10 and 40) based on staining for the stromal marker CD29 and the absence of hematopoietic markers CD45, <t>CD34,</t> and the fibroblast marker CD90. Quadrant gates were set based on unstained controls. Scale bars = 100 µm.
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Basic Characterization of the IMG-A1 Granulosa Cell Line. ( a , b ) Phase-contrast micrographs of the IMG-A1 cell culture. ( c ) Staining for senescence-associated β-galactosidase activity in passage 18 cells. ( d – f ) Immunocytochemical staining for the proliferation marker Ki67, showing a phase-contrast image ( d ), Ki67 immunofluorescence ( e ), and a merged image with DAPI nuclear counterstain ( f ). ( g , h ) Representative metaphase spreads showing a normal diploid karyotype (40 chromosomes) ( g ) and a near-tetraploid karyotype (78 chromosomes) from the IMG-A1 line ( h ). ( i – k ) Ploidy analysis by flow cytometry. Control blood cells ( i ) and primary granulosa cells ( j ) show characteristic diploid (2n) and tetraploid (4n, G2/M phase) peaks. The IMG-A1 culture ( k ) displays a predominantly near-tetraploid and near-octaploid cell population. ( l ) Flow cytometry analysis confirming the homogeneity of the IMG-A1 culture (passages 10 and 40) based on staining for the stromal marker CD29 and the absence of hematopoietic markers CD45, CD34, and the fibroblast marker CD90. Quadrant gates were set based on unstained controls. Scale bars = 100 µm.

Journal: Cells

Article Title: IMG-A1: A Novel Immortalized Granulosa Cell Line for Investigating FSH-Dependent Folliculogenesis and Ovarian Pathophysiology

doi: 10.3390/cells14241940

Figure Lengend Snippet: Basic Characterization of the IMG-A1 Granulosa Cell Line. ( a , b ) Phase-contrast micrographs of the IMG-A1 cell culture. ( c ) Staining for senescence-associated β-galactosidase activity in passage 18 cells. ( d – f ) Immunocytochemical staining for the proliferation marker Ki67, showing a phase-contrast image ( d ), Ki67 immunofluorescence ( e ), and a merged image with DAPI nuclear counterstain ( f ). ( g , h ) Representative metaphase spreads showing a normal diploid karyotype (40 chromosomes) ( g ) and a near-tetraploid karyotype (78 chromosomes) from the IMG-A1 line ( h ). ( i – k ) Ploidy analysis by flow cytometry. Control blood cells ( i ) and primary granulosa cells ( j ) show characteristic diploid (2n) and tetraploid (4n, G2/M phase) peaks. The IMG-A1 culture ( k ) displays a predominantly near-tetraploid and near-octaploid cell population. ( l ) Flow cytometry analysis confirming the homogeneity of the IMG-A1 culture (passages 10 and 40) based on staining for the stromal marker CD29 and the absence of hematopoietic markers CD45, CD34, and the fibroblast marker CD90. Quadrant gates were set based on unstained controls. Scale bars = 100 µm.

Article Snippet: PE CD34 Rat mAb [RAM34] , Elabscience Biotechnology Co., Ltd., Wuhan, China , E-AB-F1284D.

Techniques: Cell Culture, Staining, Activity Assay, Marker, Immunofluorescence, Flow Cytometry, Control