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Plasminogen activator (urokinase) from cultured cells
Thrombosis and Haemostasis
|October 31, 1979
Summary
Cultured renal cells can produce high concentrations of urokinase (UK), a valuable therapeutic enzyme. Optimizing cell culture conditions, including glycine supplementation, significantly enhances UK yield for potential large-scale production.
Area of Science:
- Biotechnology
- Cell Biology
- Biochemistry
Background:
- Urokinase (UK) is a crucial enzyme with significant therapeutic applications.
- Existing methods for UK production face challenges in meeting demand and optimizing yield.
- Renal cells in culture show potential for high-volume UK secretion.
Purpose of the Study:
- To investigate the feasibility of using cultured renal cells for large-scale urokinase production.
- To identify key factors influencing urokinase secretion rates and overall yields.
- To optimize cell culture conditions for maximizing urokinase concentration.
Main Methods:
- Culturing renal cells from various species.
- Evaluating the impact of cell passage number and inoculum density on UK secretion.
- Assessing the effect of maintenance media composition, including glycine supplementation.
- Quantifying urokinase concentration using CTA units.
Main Results:
- Significant variations in urokinase secretion were observed across different species and cell passage numbers.
- Optimized culture conditions, particularly with glycine, led to urokinase concentrations around 800 CTA units/ml.
- This represents a 100-fold increase compared to concentrations found in human urine.
Conclusions:
- Tissue culture of renal cells is a viable and effective method for high-yield urokinase production.
- Glycine supplementation in maintenance media is a key factor in achieving high urokinase concentrations.
- This optimized cell culture approach offers a promising alternative for meeting the demand for therapeutic urokinase.