A novel method to isolate odontoblasts from rat incisor.
Historically, odontoblasts have been isolated from rat incisor using a surgical curette to separate these cells from the dentin. Isolation of odontoblasts using this approach typically resulted in cells with membrane properties that made the application of patch-clamp electrophysiological techniques prohibitive. The studies here describe a new procedure for isolating mature odontoblasts from adult rat incisor to obtain enriched populations of intact, viable odontoblasts that can be readily studied using patch-clamp methodologies. Identification of isolated cells as odontoblasts was confirmed using in situ mRNA hybridization for expression of dentin sialoprotein, osteocalcin, bone sialoprotein, and type I collagen, and calcium flux was monitored in these cells by means of fura-2 microfluorometry. We suggest that either single odontoblasts or clusters of these cells isolated by this new method would be an ideal preparation for the study of odontoblast properties using electrophysiological techniques, in situ hybridization and/or microfluorometry.
Guo
L
L
Division of Basic Sciences, New York University College of Dentistry, 345 East 24th Street, New York, New York 10010, USA.
Berry
J E
JE
Somerman
M J
MJ
Davidson
R M
RM
D000818:Animals; D048429:Cell Size; D003094:Collagen / Q000235:genetics; D017364:Collagenases / Q000494:pharmacology; D003592:Cytophotometry; D003851:Deoxyribonucleases / Q000494:pharmacology; D004594:Electrophysiology; D016326:Extracellular Matrix Proteins; D005260:Female; D017403:In Situ Hybridization; D007180:Incisor / Q000166:cytology*; D008297:Male; D008722:Methods; D009804:Odontoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015675:Osteocalcin / Q000235:genetics; D010196:Pancreatic Elastase / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010750:Phosphoproteins; D011498:Protein Precursors; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012795:Sialoglycoproteins / Q000235:genetics; D000099311:Dentin Sialophosphoprotein
D003094:Collagen; D017364:Collagenases; D003851:Deoxyribonucleases; D016326:Extracellular Matrix Proteins; D015675:Osteocalcin; D010196:Pancreatic Elastase; D010750:Phosphoproteins; D011498:Protein Precursors; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D000099311:Dentin Sialophosphoprotein