title: Brain stem slice conditioned medium contains endogenous BDNF and GDNF that affect neural crest boundary cap cells in co-culture.
abstract: Conditioned medium (CM), made by collecting medium after a few days in cell culture and then re-using it to further stimulate other cells, is a known experimental concept since the 1950s. Our group has explored this technique to stimulate the performance of cells in culture in general, and to evaluate stem- and progenitor cell aptitude for auditory nerve repair enhancement in particular. As compared to other mediums, all primary endpoints in our published experimental settings have weighed in favor of conditioned culture medium, where we have shown that conditioned culture medium has a stimulatory effect on cell survival. In order to explore the reasons for this improved survival we set out to analyze the conditioned culture medium. We utilized ELISA kits to investigate whether brain stem (BS) slice CM contains any significant amounts of brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF). We further looked for a donor cell with progenitor characteristics that would be receptive to BDNF and GDNF. We chose the well-documented boundary cap (BC) progenitor cells to be tested in our in vitro co-culture setting together with cochlear nucleus (CN) of the BS. The results show that BS CM contains BDNF and GDNF and that survival of BC cells, as well as BC cell differentiation into neurons, were enhanced when BS CM were used. Altogether, we conclude that BC cells transplanted into a BDNF and GDNF rich environment could be suitable for treatment of a traumatized or degenerated auditory nerve.
authors.affiliation: Department of Clinical Science, Intervention and Technology, Karolinska University Hospital, 17176 Stockholm, Sweden. Electronic address: andreas.kaiser@karolinska.se.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kaiser
authors.affiliation: Department of Clinical Science, Intervention and Technology, Karolinska University Hospital, 17176 Stockholm, Sweden.
authors.forename: Ajay
authors.initials: A
authors.lastname: Kale
authors.affiliation: Department of Clinical Science, Intervention and Technology, Karolinska University Hospital, 17176 Stockholm, Sweden.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Novozhilova
authors.affiliation: Department of Clinical Science, Intervention and Technology, Karolinska University Hospital, 17176 Stockholm, Sweden.
authors.forename: Piyaporn
authors.initials: P
authors.lastname: Siratirakun
authors.affiliation: Unit of Molecular Neurobiology-MBB, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Jorge B
authors.initials: JB
authors.lastname: Aquino
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Charoensri
authors.initials: C
authors.lastname: Thonabulsombat
authors.affiliation: Unit of Molecular Neurobiology-MBB, Karolinska Institutet, 17177 Stockholm, Sweden.
authors.forename: Patrik
authors.initials: P
authors.lastname: Ernfors
authors.affiliation: Department of Clinical and Experimental Medicine, Faculty of Health Sciences, 58183 Linköping, Sweden.
authors.forename: Petri
authors.initials: P
authors.lastname: Olivius
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000196:embryology / Q000378:metabolism*; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism*; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism*; D051379:Mice; D009432:Neural Crest / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D017077:Culture Media, Conditioned; D051100:Glial Cell Line-Derived Neurotrophic Factor
keywords: Auditory brainstem slice; Boundary cap cells; Hearing; Neural crest; Neuronal restoration; Organotypic culture