Simone A A Romariz1, Daisyléa S Paiva1, Layla T Galindo2, Gabriela F Barnabé3, Vivian A Guedes4, Cesario V Borlongan4, Beatriz M Longo1. 1. Departamento de Fisiologia, Universidade Federal de São Paulo-UNIFESP, São Paulo, SP, Brazil. 2. Departamento de Bioquímica, Universidade Federal de São Paulo-UNIFESP, São Paulo, SP, Brazil. 3. Ludwig Institute for Cancer Research at Instituto Sírio-Libanês de Ensino e Pesquisa, São Paulo, SP, Brazil. 4. Department of Neurosurgery and Brain Repair, University of South Florida, Tampa, FL, USA.
Abstract
AIMS: Medial ganglionic eminence (MGE) progenitors give rise to inhibitory interneurons and may serve as an alternative cell source for large-scale cell transplantation for epilepsy after in vitro expansion. We investigated whether modifications in the culture medium of MGE neurospheres affect neuronal differentiation and expression of MGE-specific genes. In vivo, we compared anticonvulsant effects and cell differentiation pattern among neurospheres grown in different culture media and compared them with freshly harvested MGE cells. METHODS: We used four variations of cell culture: standard, containing growth factors (EGF/FGF-2) (GF); addition of retinoic acid (GF-RA); withdrawal of EGF/FGF-2 (WD); and addition of retinoic acid and withdrawal of EGF/FGF-2 (WD-RA). Based on in vitro results neurosphere-grown (WD-RA or GF conditions) or fresh MGE cells were transplanted into the hippocampus. RESULTS: In vitro WD-RA showed increased neuronal population and higher expression of Dlx1, Nkx2.1, and Lhx6 genes in comparison with GF culture condition. After transplantation, fresh MGE cells and neurospheres (GF) showed anticonvulsant effects. However, fresh MGE cells differentiated preferentially into inhibitory neurons, while GF gave rise to glial cells. CONCLUSION: We conclude that freshly isolated and neurosphere-grown MGE cells reduced seizures by different mechanisms (inhibitory interneurons vs. astrocytes). Fresh MGE cells appear more appropriate for cell therapies targeting inhibitory interneurons for conferring anticonvulsant outcomes.
AIMS: Medial ganglionic eminence (MGE) progenitors give rise to inhibitory interneurons and may serve as an alternative cell source for large-scale cell transplantation for epilepsy after in vitro expansion. We investigated whether modifications in the culture medium of MGE neurospheres affect neuronal differentiation and expression of MGE-specific genes. In vivo, we compared anticonvulsant effects and cell differentiation pattern among neurospheres grown in different culture media and compared them with freshly harvested MGE cells. METHODS: We used four variations of cell culture: standard, containing growth factors (EGF/FGF-2) (GF); addition of retinoic acid (GF-RA); withdrawal of EGF/FGF-2 (WD); and addition of retinoic acid and withdrawal of EGF/FGF-2 (WD-RA). Based on in vitro results neurosphere-grown (WD-RA or GF conditions) or fresh MGE cells were transplanted into the hippocampus. RESULTS: In vitro WD-RA showed increased neuronal population and higher expression of Dlx1, Nkx2.1, and Lhx6 genes in comparison with GF culture condition. After transplantation, fresh MGE cells and neurospheres (GF) showed anticonvulsant effects. However, fresh MGE cells differentiated preferentially into inhibitory neurons, while GF gave rise to glial cells. CONCLUSION: We conclude that freshly isolated and neurosphere-grown MGE cells reduced seizures by different mechanisms (inhibitory interneurons vs. astrocytes). Fresh MGE cells appear more appropriate for cell therapies targeting inhibitory interneurons for conferring anticonvulsant outcomes.
Authors: T Ostenfeld; Etienne Joly; Yu Tzu Tai; Anna Peters; Maeve Caldwell; Eric Jauniaux; C N Svendsen Journal: Brain Res Dev Brain Res Date: 2002-03-31
Authors: Claire M Kelly; Pam Tyers; Melanie Ter Borg; Clive N Svendsen; Stephen B Dunnett; Anne E Rosser Journal: Brain Res Bull Date: 2005-09-21 Impact factor: 4.077
Authors: Manuel Alvarez-Dolado; Maria Elisa Calcagnotto; Kameel M Karkar; Derek G Southwell; Dorothy M Jones-Davis; Rosanne C Estrada; John L R Rubenstein; Arturo Alvarez-Buylla; Scott C Baraban Journal: J Neurosci Date: 2006-07-12 Impact factor: 6.167
Authors: Laura da Silva Siqueira; Fernanda Majolo; Ana Paula Bornes da Silva; Jaderson Costa da Costa; Daniel Rodrigo Marinowic Journal: Mol Biol Rep Date: 2021-03-25 Impact factor: 2.316
Authors: Fu-Chen Yang; Jay L Vivian; Catherine Traxler; Steven M Shapiro; John A Stanford Journal: Cell Transplant Date: 2022 Jan-Dec Impact factor: 4.139