Isolation of specific bone cell types by free-flow electrophoresis.
The bone cells and fibroblasts from fetal rat calvaria can be isolated by collagenase digestion of the collagen matrix and separated into specific cell types by free-flow electrophoresis. The method involves injection of a specially prepared suspension of cells into a stream of buffer across which is maintained an electric field of 60 V/cm. The fetal bone cell types are differentially deflected toward the anode where they can be collected. Free-flow electrophoresis of this heterogenous cell preparation yields three distinguishable peaks which can be identified by morphologic, morphometric, and enzymatic characteristics. All three cell peaks have greater than 95% viability as judged by trypan blue exclusion and will grow to confluent monolayers in culture. The data indicate that these cell peaks may be comprised of osteoclasts and/or preosteoclasts, osteoblasts and/or preosteoblasts, and fibroblasts.
D000086:Acetazolamide / Q000378:metabolism; D000135:Acid Phosphatase / Q000032:analysis; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001842:Bone and Bones / Q000166:cytology* / Q000378:metabolism; D002469:Cell Separation; D004247:DNA / Q000096:biosynthesis; D004586:Electrophoresis / Q000379:methods; D005333:Fetus; D051381:Rats
D004247:DNA; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D000086:Acetazolamide