Comparison of P19-Derived Neuroprogenitor and Naive Cell Survival after Intracerebellar Application into B6CBA Mice

Houdek, Z.; Cendelín, J.; Kulda, V.; Babuška, V.; Vožeh, F.; Hatina, J.; Králíčková, Milena; Zech, N. H.; Veselá, I.; Pacherník, J.; Uher, P.

doi:10.14712/fb2011057040162

Folia Biologica > Archive > 2011, Vol. 57 > Issue 4 > Comparison of P19-Derived…

Fol. Biol. 2011, 57, 162-169

https://doi.org/10.14712/fb2011057040162

Comparison of P19-Derived Neuroprogenitor and Naive Cell Survival after Intracerebellar Application into B6CBA Mice

Z. Houdek^1,2, J. Cendelín¹, V. Kulda³, V. Babuška³, F. Vožeh¹, J. Hatina⁴, Milena Králíčková^2,5, N. H. Zech⁶, I. Veselá⁷, J. Pacherník⁷, P. Uher²

¹Department of Pathophysiology, Charles University in Prague – Faculty of Medicine in Pilsen, Pilsen, Czech Republic
²IVF Centers Prof. Zech – Pilsen s. r. o., Pilsen, Czech Republic
³Department of Medical Chemistry and Biochemistry, Charles University in Prague – Faculty of Medicine in Pilsen, Pilsen, Czech Republic
⁴Department of Biology, Charles University in Prague – Faculty of Medicine in Pilsen, Pilsen, Czech Republic
⁵Department of Histology and Embryology Charles University in Prague – Faculty of Medicine in Pilsen, Pilsen, Czech Republic
⁶IVF Centers Prof. Zech – Bregenz, Bregenz, and Department for Obstetrics and Gynecology, Unit of Gynecological Endocrinology and Reproductive Medicine, University of Graz, Austria
⁷Department of Animal Physiology and Immunology, Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic

Received March 2011
Accepted March 2011

Mouse embryonic carcinoma cells (P19 line) were studied for both their survival and developmental potential in the intact cerebellum of B6CBA mice. The P19 cells were cultured and labelled with green fluorescent protein using transfection. Cells were used for transplantation either in the undifferentiated stage or after 3 days of neurodifferentiation induced by retinoic acid. The intracerebellar application was performed in 43 mice: group A (N = 21) received neuroprogenitors and group B (N = 22) received undifferentiated cells. The morphology of transplanted cells within the context of the surrounding cerebellar tissue was evaluated after 3 weeks. Naive P19 cells engrafted and survived in the cerebellum of 7 of the 22 adult mice (survival rate 31.8 %). Neuroprogenitors survived in 13 of the 21 mice (survival rate was 61.9 %). Since the cut-off is P < 0.05, the difference is not statistically significant (P = 0.069). An expansive appearance of the graft was significantly more frequent (P = 0.0047) in naive P19 cells than in neuroprogenitors. In mice in which the grafts did not survive, no marks of grafted cells or only fluorescing detritus were found. In conclusion, this is the first study to track the fate and morphology of embryonic carcinoma cells transplanted into the cerebellum, confirming that neuroprogenitors derived from embryonic carcinoma cells can settle in the host tissue and differentiate according to the surrounding conditions. With further validation, the embryonic carcinoma cells could become a valuable model with which to study the impact of cell therapy on neurodegenerative diseases.

Keywords

P19-derived neuroprogenitors, naive P19 cells, intracerebellar application, graft survival.

Funding

This study was supported by research project of the Ministry of Education, Youth and Sports of the Czech Republic VZ MSM 021620816, grant COST BM0603 No. OC10038 of the Ministry of Education, Youth and Sports of the Czech Republic, grant from the Czech Science Foundation No. 301/08/0717, and by specific student research grant of Charles University SVV-2011-262 806.