A new approach for the characterization of proliferative cells in cestodes.
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.
Sección Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.
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MF
Domínguez
Sección Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay; Institute for Hygiene and Microbiology, University of Würzburg, Würzburg, Germany.
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U
Koziol
Unidad de Biología Celular, Institut Pasteur de Montevideo, Montevideo, Uruguay.
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Porro
Sección Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay; Departamento de Genética, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
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A
Costábile
Sección Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.
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Estrade
Departamento de Genética, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
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J
Tort
Unidad de Biología Celular, Institut Pasteur de Montevideo, Montevideo, Uruguay.
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M
Bollati-Fogolin
Sección Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. Electronic address: estelacasti@gmail.com.
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E
Castillo
D000818:Animals; D001562:Benzimidazoles; D002453:Cell Cycle; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D007814:Larva / Q000166:cytology; D008645:Mesocestoides / Q000166:cytology* / Q000254:growth & development; D051379:Mice; D023421:Models, Animal; D011419:Propidium; D015203:Reproducibility of Results; D014357:Trypsin / Q000378:metabolism
D001562:Benzimidazoles; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; C085925:calcein AM; D011419:Propidium; D014357:Trypsin; C017807:bisbenzimide ethoxide trihydrochloride
Cestodes; FACS; Flow cytometry; Mesocestoides corti; Proliferative cells