title: In vitro culture of neurons from sheep with Batten disease.
abstract: Specific storage of mitochondrial ATP synthase subunit c occurs in most forms of Batten disease, including the ovine form, but its relationship to the characteristic neurodegeneration is not clear. Storage occurs in most cell types but only neurons are functionally affected. Neurons were cultured from control and affected sheep. Ewes were superovulated and inseminated, and embryos were collected, frozen, stored, and later transplanted into surrogate dams for gestation at times to suit experimental demands. The optimal fetal age for cultures was investigated, from 50 to 125 days. There were no differences between control and affected embryos in this period of rapid growth. At 50 days brains consist of smooth-surfaced hemispheres and cerebellum with no obvious demarcation between gray and white matter. At 90 days they are like miniature adult brains. From 200 to 600 million viable cells were recovered from each fetus, regardless of age. DMEM/F12 with B27 was the most practical medium tested. Cell viability was not as good in medium containing serum. Treatment of surfaces with polylysine aided neuron adhesion. No developmental or viability differences were observed between normal and affected neuron cultures. At plating out cells were rounded. A day later single process outgrowths began. After 4 days these were over 200 microm and by Day 6 had created a network. Most neurons were bipolar. Neurons from 50 to 90-day old fetuses persisted in culture for over 100 days.
mesh_terms: D000818:Animals; D001921:Brain / Q000196:embryology / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D005865:Gestational Age; D007150:Immunohistochemistry; D008297:Male; D009472:Neuronal Ceroid-Lipofuscinoses / Q000196:embryology / Q000378:metabolism / Q000473:pathology*; D009474:Neurons / Q000473:pathology*; D012756:Sheep