Cell separation by velocity sedimentation of postnatal mouse cerebellum

✍ Scribed by David S. Barkley; Ljiljana L. Rakic; Jonathan K. Chaffee; Dona L. Wong

• Book ID: 102881231
• Publisher: John Wiley and Sons
• Year: 1973
• Tongue: English
• Weight: 886 KB
• Volume: 81
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1040810215

✦ Synopsis

Abstract

Trypsinized cells of newborn mouse cerebellum have been separated by velocity sedimentation at unit gravity in shallow gradients of Ficoll. The two main technical difficulties were formation of gels around the dissociated cells and clumping of cells before and during the sedimentation procedure. These were solved by adding DNase to the dissociation medium and with holding serum, respectively. Proliferating cells of the external granular layer separated according to size differences in the cell generation cycle. Identification of Purkinje or other early‐forming neurons was made by labeling them with ^3^H‐thymidine on their birthdays. Many of the fractions contain viable cells capable of aggregating in culture.