Cryopreservation and primary culture of cerebral neurons from cynomolgus monkeys (Macaca fascicularis).
We established the procedures for cryopreservation and primary culture of fetal cerebral neurons of cynomolgus monkeys (Macaca fascicularis). Three developmental stages of fetuses (80, 93, and 102 days of gestation) were compared to determine the optimal stage of cerebrum development for primary culture. Among the three fetuses, the 80-day-old fetus produced the most process-rich neurons with the highest survival. The number of total recovery cells from the cryopreserved 80-day-old fetus corresponded to 83.4% of that from fresh tissue. Besides, synchronous oscillations of intracellular calcium were first seen in primate cerebral neurons, which suggested the formation of synapse-networks. Cultured neurons expressed synaptophysin protein. Successful cryopreservation and subsequent cell culture of primate neurons would be useful tools for neuroscience research with species specificity.
Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. taka-u@yayoi.club.ne.jp
Takayuki
T
Negishi
Yoshiyuki
Y
Ishii
Seiji
S
Kawamura
Yoichiro
Y
Kuroda
Yasuhiro
Y
Yoshikawa
D000818:Animals; D001683:Biological Clocks / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured / Q000166:cytology* / Q000378:metabolism; D002540:Cerebral Cortex / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D005260:Female; D005333:Fetus / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D008252:Macaca fascicularis / Q000196:embryology* / Q000378:metabolism / Q000601:surgery; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009415:Nerve Net / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy; D013569:Synapses / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology; D016708:Synaptophysin / Q000378:metabolism
D003451:Cryoprotective Agents; D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D016708:Synaptophysin; D002118:Calcium