Category Archives: Fibroblast

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: DISCUSSION(4)

We also tested the response of fetal fibroblasts to cell cycle inhibitors DMSO, mimosine, and colchicine, which have been shown to arrest cultured cells in G0, G1, and G2+M, respectively. Addition of 1% DMSO to cycling cultures for 48 h … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: DISCUSSION(3)

By gating on small-, medium-, and large-sized cells, we found that percentages of cells existing in the various phases of the cell cycle differed due to cell size. As the cell size decreased from large to small, percentages of cells … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: DISCUSSION(2)

It is known that, for certain cell types, serum starvation is not a viable method for synchronizing cells into G0, either because the cells arrest in G1 instead or because they undergo apoptosis. Since quiescent cells contain characteristically low levels … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: DISCUSSION(1)

The development of reconstructed embryos following nuclear transfer appears to be dependent upon a variety of factors. The most important factor identified thus far is cell cycle synchrony of donor nuclei with recipient enucleated oocytes (reviewed in ). Donor nuclei … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: RESULTS(3)

Based on forward light scatter, a measurement of particle (cell) size, percentages of cells in the various stages of the┬ácell cycle were calculated when gates were set to include only small-, medium-, or large-sized cells (Fig. 4). Microscopic measurement of … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: RESULTS(2)

Scatterplots of protein versus DNA and single-parameter histograms of protein both clearly show a marked decline in the levels of cellular protein of fetal cells as a consequence of serum deprivation (Fig. 2). As made apparent by the wide-spreading green … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: RESULTS(1)

Figure 1 represents the distribution of cycling fetal cells existing in the various phases of the cell cycle as determined by flow cytometry. Single-parameter histograms of DNA (red fluorescence) provided data for percentages of cells existing in G0+G1 (2C DNA … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: MATERIALS AND METHODS(4)

CellTreatments Cell cycle comparisons were made between cycling cells, serum-starved cells, and cells cultured to confluency. Flasks were seeded with fetal cells (passages 5-10) at a concentration of 105 cells/ml. After 2 days of culture, cells were allocated to one … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: MATERIALS AND METHODS(3)

Flow Cytometry and Confocal Microscopy Stained cells, suspended in the staining solution, were analyzed using a Becton Dickinson (Rutherford, NJ) flow cytometer as described by Crissman and Steinkamp. Fluorescence data were collected using 488-nm excitation and filter combination consisting of … Continue reading

Flow Cytometric Cell Cycle Analysis of Cultured Porcine: MATERIALS AND METHODS(2)

Cell Fixation and Staining Fetal cells were analyzed for protein and DNA content by ethanol fixation and staining with propidium iodide and fluorescein isothiocyanate using a modified method described by Crissman and Steinkamp. Trypsinized cells were resuspended in DMEM with … Continue reading