A new method of embryonic culture for assessing global changes in brain organization.
While dissociated, reaggregated cells and organotypic slice cultures are useful models for understanding brain development, they only partially mimic the processes and organization that exist in vivo. Towards bridging the gap between in vitro and in vivo paradigms, a method for culturing intact brain tissue was developed using whole cerebral cortical hemispheres in which the anatomical and cellular organization of nervous system tissue is preserved. Single, free-floating telencephalic hemispheres were dissected from embryonic mice and placed into defined culture medium on an orbital shaker. Orbital shaking was necessary for optimal growth, and cortices grown under these conditions closely approximated in vivo parameters of cell division, differentiation, migration and cell death for up to 24 h. In addition to wild-type cultures, the method was compatible with genetically altered tissues. One particular advantage of this method is its ability to reveal global anatomical alterations in the embryonic brain following exposure to soluble growth factors. This method should thus be helpful for assessing a wide range of soluble molecules for their systemic effects on the embryonic brain.
Rehen
Stevens K
SK
Helen L. Dorris Child and Adolescent Neuropsychiatric Disorder Institute, The Scripps Research Institute, La Jolla, CA 92037, USA.
Kingsbury
Marcy A
MA
Almeida
Beatriz S V
BS
Herr
Deron R
DR
Peterson
Suzanne
S
Chun
Jerold
J
D000818:Animals; D001921:Brain / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000166:cytology* / Q000502:physiology; D005260:Female; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000502:physiology; D011247:Pregnancy; D013997:Time Factors
D009419:Nerve Tissue Proteins; D053148:Caspase 3