Calcified Tissue International

, Volume 27, Issue 1, pp 263–268

Isolation of specific bone cell types by free-flow electrophoresis

  • J. Edward Puzas
  • Agnes Vignery
  • Howard Rasmussen
Laboratory Investigations
  • 29 Downloads

Summary

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.

Key words

Bone cells Separation Free-flow electrophoresis 

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Copyright information

© Springer-Verlag 1979

Authors and Affiliations

  • J. Edward Puzas
    • 1
  • Agnes Vignery
    • 1
  • Howard Rasmussen
    • 1
  1. 1.Departments of Internal Medicine and PathologyYale University School of MedicineNew HavenUSA
  2. 2.V.A. Medical Center, Research Service (11A), American LakeTacomaUSA

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