| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 4371160 | Experimental Parasitology | 2014 | 5 Pages |
•Optimization of a method to obtain cell suspensions of the cestode Mesocestoides corti.•Cell cycle analysis of cell suspensions by flow cytometry.•Isolation of cellular fractions in G0/G1 and G2/M phase of M. corti by cell sorting.•The cells isolated on the G2/M fraction have the characteristics of germinative cells.•It is possible to obtain RNA from the cells isolated by FACS.
Cestodes show a remarkable proliferative capability that sustains the constant growth and differentiation of proglottids essential for their lifestyle. It is believed that a separate population of undifferentiated stem cells (the so-called germinative cells) are the only cells capable of proliferation during growth and development. The study of this particular cell subpopulation is hampered by the current lack of methods to isolate it. In this work, we developed a reproducible flow cytometry and cell sorting method to quantify and isolate the proliferating cells in the tetrathyridia larvae of the model cestode Mesocestoides corti, based on the DNA content of the cells. The isolated cells display the typical germinative cell morphology, and can be used for RNA isolation with a yield in the ng to μg range. We expect that this approach may facilitate the characterization of the germinative cells in M. corti and other model tapeworms.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slide
