Studies of the metabolism of separated bone cells

Smith, Duane M.; Johnston, C. Conrad; Severson, Arlen R.

doi:10.1007/bf02546595

Cited by 45 publications

(6 citation statements)

References 25 publications

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“…Histochemical analysis of cells obtained from the third passage of culture six was employed to detect the presence of alkaline phosphatase which characterizes osteoblasts and some osteocytes [10], and acid phosphatase [11] and succinic dehydrogenase which characterize bone resorbing cells [12]. Cells grown on coverslips were washed with PBS, pH 7.4, and incubated for 30 min at 37~ with Sigma naphthol AS-MX phosphate, pH 8.4, containing Fast Blue RR to identify alkaline phosphatase activity.…”

Section: Histochemical Proceduresmentioning

confidence: 99%

Long-term culture of cells from bone affected by Paget’s disease

et al. 1979

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Cells obtained from surgical bone specimens of eight patients with Paget's disease of bone were maintained in culture for up to 8 months and seven passages. The doubling time during the period of maximal cell growth ranged from 4 to 12 days. Evidence consistent with the hypothesis that many of the cells were bone cells included the following: (a) histochemical techniques demonstrated staining of some cells for alkaline phosphatase or acid phosphatase and succinic dehydrogenase; (b) parathyroid extract stimulated increased uptake of 3H-thymidine and 3H-uridine; (c) parathyroid extract suppressed and salmon calcitonin stimulated uptake of 3H-proline; and (d) crystalline calcium deposits were found within cells and extracellularly. Ultrastructural analysis revealed that three of the eight cultures contained cells whose nuclei had inclusions which were almost identical to those found in the osteoclast nuclei of all patients with Paget's disease. The maintenance of cells derived from pagetic bone in long-term culture should aid in testing the hypothesis that Paget's disease represents a slow virus infection of bone.

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Section: Histochemical Proceduresmentioning

confidence: 99%

Long-term culture of cells from bone affected by Paget’s disease

et al. 1979

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“…The preparation of diaphysis, however, corresponded to the osteocytic tissue preparation of Smith et al(1973), and was (Exp I). The absence of a reciprocal relationship between cyclic GMP and cyclic AMP was seen also in experiment in which PTH or calcitonin, either at low or high concentration, similarly elevated cyclic AMP levels in the presence of carbachol as in its absence (Exp.…”

Section: Resultsmentioning

confidence: 99%

“…The fact that adrenergic and cholinergic transmitters both affect the metabolism of cyclic nucleotides in bone, as shown in the present study, may be regarded as evidence that the activities of bone cells can be directly influenced by autonomic nerves. The response detected in the preparation of diaphysis which corresponded to the osteocytic tissue preparation of Smith et al(1973) probably indicates a direct involvement in the response of bone cells rather than contaminants such as marrow cells (Table 1).…”

Section: Discussionmentioning

confidence: 99%

Effects of adrenergic and cholinergic stimulations on cyclic nucleotides metabolism in bone of young rat.

1978

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Epinephrine or carbachol, alone or together with parathyroid hormone (PTH) or calcitonin, was added into the incubation of young rat tibia and cyclic AMP as well as cyclic GMP in the tissue was determined. Epinephrine induced a dose related and rapid increase of cyclic AMP probably through mechanism different from that of PTH or calcitonin. The effect of these hormones to increase cyclic AMP was additive to the effect of maximal dose of epinephrine and, in contrary to epinephrine effect, was not influenced by propranolol. Carbachol induced a dose-related and rapid increase of cyclic GMP in bone. The effect of carbachol was completely abolished by adding atropine or tetracaine. A marked increase of cyclic GMP was also induced by NaN3. There was no reciprocal relationship between the levels of cyclic AMP and cyclic GMP. The increase of cyclic GMP by carbachol was unaffected by the increase of cyclic AMP induced by PTH, and the increase of cyclic AMP by PTH or calcitonin was the same in the presence or absence of carbachol at the dose maximally effective to increase cyclic GMP. The results indicate the presence in bone of adrenergic and cholinergic receptor sites, and possible regulation of bone cell activity by these nervous systems through cyclic AMP and cyclic GMP. Though the regulation of the cyclic nucleotides content in bone by these neurotransmitters appeared to be through mechanism independent of PTH or calcitonin action, it seems worthwhile to study further how these neurotransmitters by themselves or in concert with these hormones act on the various aspects of bone cell function.

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“…However, a limitation in their use in studies of metabolic control is the fact that the cells prepared by most present isolation methods are a heterogenous population, composed of many cell types. Several methods have been described for solving this problem and obtaining populations of isolated bone cell types [6][7][8][9]. Rat or mouse fetal calvaria have been used as starting material and the separation techniques have involved manual dissection and/or sequential digestion of the tissue by bacterial collagenase.…”

Section: Introductionmentioning

confidence: 99%

Isolation of specific bone cell types by free-flow electrophoresis

Puzas¹,

Vignery²,

Rasmussen³

1979

Calcif Tissue Int

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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.

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Studies of the metabolism of separated bone cells

Cited by 45 publications

References 25 publications

Long-term culture of cells from bone affected by Paget’s disease

Long-term culture of cells from bone affected by Paget’s disease

Effects of adrenergic and cholinergic stimulations on cyclic nucleotides metabolism in bone of young rat.

Isolation of specific bone cell types by free-flow electrophoresis

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