title: Survival, migration, and differentiation of Sox1-GFP embryonic stem cells in coculture with an auditory brainstem slice preparation.
abstract: The poor regeneration capability of the mammalian hearing organ has initiated different approaches to enhance its functionality after injury. To evaluate a potential neuronal repair paradigm in the inner ear and cochlear nerve we have previously used embryonic neuronal tissue and stem cells for implantation in vivo and in vitro. At present, we have used in vitro techniques to study the survival and differentiation of Sox1-green fluorescent protein (GFP) mouse embryonic stem (ES) cells as a monoculture or as a coculture with rat auditory brainstem slices. For the coculture, 300 microm-thick brainstem slices encompassing the cochlear nucleus and cochlear nerve were prepared from postnatal SD rats. The slices were propagated using the membrane interface method and the cochlear nuclei were prelabeled with DiI. After some days in culture a suspension of Sox1 cells was deposited next to the brainstem slice. Following deposition Sox1 cells migrated toward the brainstem and onto the cochlear nucleus. GFP was not detectable in undifferentiated ES cells but became evident during neural differentiation. Up to 2 weeks after transplantation the cocultures were fixed. The undifferentiated cells were evaluated with antibodies against progenitor cells whereas the differentiated cells were determined with neuronal and glial markers. The morphological and immunohistochemical data indicated that Sox1 cells in monoculture differentiated into a higher percentage of glial cells than neurons. However, when a coculture was used a significantly lower percentage of Sox1 cells differentiated into glial cells. The results demonstrate that a coculture of Sox1 cells and auditory brainstem present a useful model to study stem cell differentiation.
authors.affiliation: Center for Hearing and Communication Research, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Glavaski-Joksimovic
authors.forename: Charoensri
authors.initials: C
authors.lastname: Thonabulsombat
authors.forename: Malin
authors.initials: M
authors.lastname: Wendt
authors.forename: Mikael
authors.initials: M
authors.lastname: Eriksson
authors.forename: Björn
authors.initials: B
authors.lastname: Palmgren
authors.forename: Anna
authors.initials: A
authors.lastname: Jonsson
authors.forename: Petri
authors.initials: P
authors.lastname: Olivius
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001303:Auditory Cortex / Q000502:physiology*; D001933:Brain Stem / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D002470:Cell Survival; D018920:Coculture Techniques; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D005260:Female; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D055748:SOXB1 Transcription Factors
chemical_list: D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D011993:Recombinant Fusion Proteins; D055748:SOXB1 Transcription Factors; C098693:Sox1 protein, mouse; D049452:Green Fluorescent Proteins