Possibilities and limitations of bone organ culture
Possibilities and limitations of bone organ culture
The possibilities and limitations of culturing relatively mature femurs from 15-day old chick embryos was investigated. During a 6-8 day culture period the femurs sythesised alkaline phosphatase and replenished ascorbate stores by accumulation from the medium. The cartilage epiphyses expanded considerably and the formation of new osteoid was observed in histological sections. The rate of bone and cartilage formation was quantified by measuring collagen synthesis. Since culture conditions were generally more suitable for the growth of cartilage than that of bone, attempts were made to improve conditions for bone culture. Insulin, albumin and conalbumin had no effect whereas certain non-essential amino acids had considerable effects on growth and collagen synthesis. In particular glycine was essential for growth, but high concentrations of alanine inhibited [3 H]-proline uptake. By judiciously altering the concentrations of these amino acids, collagen synthesis could be increased by 100%. Uptake of 45Ca was studied to assess mineralisation. Even though 98% of incorporated 45 Ca was present in the bone shafts, this was not sufficient to indicate de novo mineralisation. β-glycerophosphate stimulated ^45Ca influx into cartilage but not into bone and prevented the synthesis of alkaline phosphatase. Fetal bovine serum stimulated proliferation but inhibited ^45 Ca uptake. The major limitations of bone organ culture were considered to be the diffusion constraints of the bone matrix, insufficient proliferation of skeletal precursor cells and absence of factors inducing mineralisation of endochondral ossification. Bone organ culture is suitable for studies related to the earliest stage of osteoid formation, the control of differentiation in bone and cartilage, the resorption of non-mineralised bone and cartilage and for the investigation of the relationship between bone resorption and bone formation.
University of Southampton
1987
Roach, Helmtrud Isolde
(1987)
Possibilities and limitations of bone organ culture.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The possibilities and limitations of culturing relatively mature femurs from 15-day old chick embryos was investigated. During a 6-8 day culture period the femurs sythesised alkaline phosphatase and replenished ascorbate stores by accumulation from the medium. The cartilage epiphyses expanded considerably and the formation of new osteoid was observed in histological sections. The rate of bone and cartilage formation was quantified by measuring collagen synthesis. Since culture conditions were generally more suitable for the growth of cartilage than that of bone, attempts were made to improve conditions for bone culture. Insulin, albumin and conalbumin had no effect whereas certain non-essential amino acids had considerable effects on growth and collagen synthesis. In particular glycine was essential for growth, but high concentrations of alanine inhibited [3 H]-proline uptake. By judiciously altering the concentrations of these amino acids, collagen synthesis could be increased by 100%. Uptake of 45Ca was studied to assess mineralisation. Even though 98% of incorporated 45 Ca was present in the bone shafts, this was not sufficient to indicate de novo mineralisation. β-glycerophosphate stimulated ^45Ca influx into cartilage but not into bone and prevented the synthesis of alkaline phosphatase. Fetal bovine serum stimulated proliferation but inhibited ^45 Ca uptake. The major limitations of bone organ culture were considered to be the diffusion constraints of the bone matrix, insufficient proliferation of skeletal precursor cells and absence of factors inducing mineralisation of endochondral ossification. Bone organ culture is suitable for studies related to the earliest stage of osteoid formation, the control of differentiation in bone and cartilage, the resorption of non-mineralised bone and cartilage and for the investigation of the relationship between bone resorption and bone formation.
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Published date: 1987
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Local EPrints ID: 461345
URI: http://eprints.soton.ac.uk/id/eprint/461345
PURE UUID: 9ca47ba5-fbe1-4dae-9625-849a014706c0
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Date deposited: 04 Jul 2022 18:43
Last modified: 04 Jul 2022 18:43
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Author:
Helmtrud Isolde Roach
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