Cortical cell elution by sedimentation field-flow fractionation.
As a cell sorter, Sedimentation field-flow fractionation (SdFFF) can be defined as an effective tool for cell separation and purification, respecting integrity and viability as well as providing enhanced recovery and purified sterile fraction collection. The complex cell suspension containing both neurons and glial cells of all types, obtained from cerebral cortices of 17-day-old rat fetuses, is routinely used as a model of primary neuronal culture. Using SdFFF, this complex cell mixture was eluted in sterile fractions which were collected and cultured. SdFFF cell elution was conducted under strictly defined conditions: rapid cell elution, high recovery (negligible cell trapping), short- and long-term cell viability, sterile collection. After immunological cellular type characterization (neurons and glial cells) of cultured cells, our results demonstrated the effectiveness of SdFFF to provide, in less than 6 min, viable and enriched neurons which can be cultured for further investigations.
Battu
S
S
Laboratoire de Chimie Analytique et Bromatologie, Faculté de Pharmacie, Université de Limoges, France. battu@pharma.unilim.fr
Elyaman
W
W
Hugon
J
J
Cardot
P J
PJ
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002540:Cerebral Cortex / Q000166:cytology* / Q000196:embryology; D005865:Gestational Age; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology*; D051381:Rats; D013194:Staining and Labeling