Calcified Tissue Research

, Volume 11, Issue 1, pp 56–69 | Cite as

Studies of the metabolism of separated bone cells

I. Techniques of separation and identification
  • D. M. Smith
  • C. C. JohnstonJr.
  • A. R. Severson
Original Papers

Abstract

The mechanical separation of rat skeletal tissue into viable populations of periosteal cells, osteoblasts, osteocytes, and marrow cells has been undertaken. Periosteal cells and osteocytes were obtained as tissue preparations, while osteoblasts and marrow cells were isolated as individual cells. The cell populations were identified during the preparative procedure by their histological and histochemical appearance. Viability of these preparations was demonstrated by their metabolic activity during short-term incubations and their histological appearance and vital staining before and after such incubations. The cell populations produced carbon dioxide and lactate, consumed oxygen, and incorporated uridine into RNA. Comparison of uridine incorporation into RNA of cells from control and parathyroid hormone treated rats was compatible with a differential effect of parathyroid hormone on the cell populations. These findings indicate that the technique described can be utilized to produce viable bone cells of different types, and may offer a new means for characterizing the role of each cell type in the metabolism of skeletal tissue.

Key words

Bone Osteoblasts Osteocytes Periosteum Marrow Cells 

Résumé

La séparation mécanique du tissu squelettique du rat en cellules périostées, en ostéoblastes, en ostéocytes et en cellules de la moelle a été réalisée. Les cellules périostées et les ostéocytes ont été obtenues sous forme de préparations tissulaires, alors que les cellules ostéoblastiques et celles de la moelle ont été isolées sous forme de cellules individuelles. Les populations cellulaires sont identifiées par leurs caractères histologiques et histochimiques. La vitalité de ces préparations est věrifiée par leur activité métabolique pendant des incubations de courte durée et des examens. histologiques et des colorations vitales, réalisés avant et après incubation. Les cellules produisent du gaz carbonique et du lactate, consomment de l’oxygène et incorporent l’uridine en RNA. La comparaison de l’incorporation d’uridine en RNA de cellules de rats témoins et de rats traités par hormone parathyroïdienne concorde avec une action différentielle de la parathormone sur les divers types de cellules. Il semble que la technique décrite permette de produire des cellules osseuses vivantes de divers types et fournit ainsi le moyen d’élucider le rôle métabolique de chaque type cellulaire du squelette.

Zusammenfassung

Die mechanische Trennung von Rattenskelett-Gewebe in lebensfähige Populationen von Periost-Zellen, Osteoblasten, Osteozyten und Markzellen ist unternommen worden. Die Periostzellen und Osteozyten wurden als Gewebepräparate erhalten, während die Osteoblasten und Markzellen als individuelle Zellen isolier wurden. Die Zell-Populationen wurden während des Herstellungsverfahrens histologisch und histochemisch identifiziert. Die Lebensfähigkeit dieser Präparate wurde festgestellt durch ihre metabolishe Aktivität während Kurzzeit-Inkubationen und durch ihr histologisches Aussehen sowie durch Vitalfärbung vor und nach solchen Inkubationen. Die Zellpopulationen produzierten Kohlendioxyd und Lactat, verbrauchten Sauerstoff und bauten Uridin in RNS ein. Der Vergleich zwischen Uridineinbau in RNS von Zellen aus Kontrolltieren und aus mit Parathormon behandelten Ratten war mit der Differentialwirkung von Parathormon auf die Zellpopulationen vereinbar. Diese Befunde deuten darauf hin, daß die beschriebene Technik benützt werden kann, um verschiedene Typen von lebensfähigen Knochenzellen zu produzieren und daß sie einen neuen Weg öffnen kann, um die Rolle jedes Zelltyps im Stoffwechsel des Skelettgewebes zu charakterisieren.

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Copyright information

© Springer-Verlag 1973

Authors and Affiliations

  • D. M. Smith
    • 3
  • C. C. JohnstonJr.
    • 1
  • A. R. Severson
    • 2
  1. 1.Division of Endocrinology and MetabolismIndiana University School of MedicineIndianapolis
  2. 2.Department of AnatomyIndiana University, School of MedicineIndianapolis
  3. 3.Department of MedicineIndiana University Medical CenterIndianapolisU.S.A.

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