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The Fibroblast Mitogenic Activity Released from Human Basophilic Cell Line KU812 is Separate from Tryptase and PDGF Expression

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The human leukaemia cell line KU812 has previously been used to study basophil differentiation. In this study the authors analysed the capacity of KU812 to produce the mast cell proteinase tryptase and to synthesize factor(s) mitogenic for fibroblasts. KU812 cells were treated with tetradecanoyl‐phorbol‐13‐acetate (TPA), conditioned medium from the human T‐cell line Mo (Mo‐CM), or cultured under serum free conditions. After 4 days the cells were analysed for cell growth, differentiation, content of tryptase, and secretion of fibroblast mitogenic activity. Mo‐CM and serum starvation increased the expression while TPA treatment down‐regulated the expression of FcεRI‐α chain. An increase in tryptase content in cell extracts was detected after 4 days of culture in serum‐free medium or in the presence of Mo‐CM. KU812 conditioned media was found to have a baseline expression of mitogenic activity on normal human foreskin fibroblasts that was increased after serum starvation or after treatment with TPA. Mast cell‐derived tryptase has previously been reported to be mitogenic for fibroblasts, but in this study the expression of tryptase did not correlate with the expression of fibroblast mitogenic activity in KU812 cells. Furthermore, affinity‐purified lung tryptase did not show any mitogenic activity. Platelet‐derived growth factor was also excluded. Although the factor(s) from KU812 cells stimulating fibroblast proliferation have not been identified, our results indicate that basophils may be potential producers of growth factors inducing fibroblast proliferation.
Title: The Fibroblast Mitogenic Activity Released from Human Basophilic Cell Line KU812 is Separate from Tryptase and PDGF Expression
Description:
The human leukaemia cell line KU812 has previously been used to study basophil differentiation.
In this study the authors analysed the capacity of KU812 to produce the mast cell proteinase tryptase and to synthesize factor(s) mitogenic for fibroblasts.
KU812 cells were treated with tetradecanoyl‐phorbol‐13‐acetate (TPA), conditioned medium from the human T‐cell line Mo (Mo‐CM), or cultured under serum free conditions.
After 4 days the cells were analysed for cell growth, differentiation, content of tryptase, and secretion of fibroblast mitogenic activity.
Mo‐CM and serum starvation increased the expression while TPA treatment down‐regulated the expression of FcεRI‐α chain.
An increase in tryptase content in cell extracts was detected after 4 days of culture in serum‐free medium or in the presence of Mo‐CM.
KU812 conditioned media was found to have a baseline expression of mitogenic activity on normal human foreskin fibroblasts that was increased after serum starvation or after treatment with TPA.
Mast cell‐derived tryptase has previously been reported to be mitogenic for fibroblasts, but in this study the expression of tryptase did not correlate with the expression of fibroblast mitogenic activity in KU812 cells.
Furthermore, affinity‐purified lung tryptase did not show any mitogenic activity.
Platelet‐derived growth factor was also excluded.
Although the factor(s) from KU812 cells stimulating fibroblast proliferation have not been identified, our results indicate that basophils may be potential producers of growth factors inducing fibroblast proliferation.

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