title: Development of a method for the purification and culture of rodent astrocytes.
abstract: The inability to purify and culture astrocytes has long hindered studies of their function. Whereas astrocyte progenitor cells can be cultured from neonatal brain, culture of mature astrocytes from postnatal brain has not been possible. Here, we report a new method to prospectively purify astrocytes by immunopanning. These astrocytes undergo apoptosis in culture, but vascular cells and HBEGF promote their survival in serum-free culture. We found that some developing astrocytes normally undergo apoptosis in vivo and that the vast majority of astrocytes contact blood vessels, suggesting that astrocytes are matched to blood vessels by competing for vascular-derived trophic factors such as HBEGF. Compared to traditional astrocyte cultures, the gene profiles of the cultured purified postnatal astrocytes much more closely resemble those of in vivo astrocytes. Although these astrocytes strongly promote synapse formation and function, they do not secrete glutamate in response to stimulation.
authors: Stanford University School of Medicine, Department of Neurobiology, Stanford, CA 94305, USA. lfoo@stanford.edu
authors: Lynette C
authors: LC
authors: Foo
authors: Nicola J
authors: NJ
authors: Allen
authors: Eric A
authors: EA
authors: Bushong
authors: P Britten
authors: PB
authors: Ventura
authors: Won-Suk
authors: WS
authors: Chung
authors: Lu
authors: L
authors: Zhou
authors: John D
authors: JD
authors: Cahoy
authors: Richard
authors: R
authors: Daneman
authors: Hui
authors: H
authors: Zong
authors: Mark H
authors: MH
authors: Ellisman
authors: Ben A
authors: BA
authors: Barres
mesh_terms: D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D001253:Astrocytes / Q000145:classification / Q000187:drug effects / Q000502:physiology*; D000067878:CELF Proteins; D002452:Cell Count / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D018925:Chemokines / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D039641:Integrin beta Chains / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000502:physiology; D062793:Occludin; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017978:Receptors, Growth Factor / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000502:physiology
chemical_list: D017304:Annexin A5; D000067878:CELF Proteins; C470022:Celf4 protein, mouse; D018925:Chemokines; D016895:Culture Media, Serum-Free; D005904:Glial Fibrillary Acidic Protein; D039641:Integrin beta Chains; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D062793:Occludin; C568986:Ocln protein, mouse; C568987:Ocln protein, rat; D016601:RNA-Binding Proteins; D017978:Receptors, Growth Factor; C084344:integrin beta5