Summary
1. Studies on uninoculated, biphasic media, containing 10% defibrinated rabbit blood in an agar base with Locke's solution as an overlay, have shown that diffusion of nutrients from the agar base into the overlay requires about 24–48 hr for equilibration.
2. Large molecules as well as nominally insoluble molecules will diffuse into the overlay and become available as nutrients.
3. The incorporation of blood into the molten agar base does not inactivate all enzymes and several active enzymes have been observed in equilibrated overlay solutions.
4. After a five day equilibration period, we have observed the following concentrations in the overlay: amino acids, 6.0 ± 1.3 mM; glucose, 146 ± 3 mg/100 ml; cholesterol, 24.8 ± 0.4 mg/100 ml; total protein, 1.50 ± 0.04 g/100 ml; albumin, 0.18 ± 0.01 g/100 ml; alkaline phosphatase, 11.5 ± 1.1 mU/ml; lactic dehydrogenase, 358 ± 20 mU/ml; and aspartic amino transferase, 4.0 ± 1.2 mU/ml.
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Stohlman, V., Mastright, G., Kazan, B.H. et al. Diffusion of Nutrients in a Biphasic Medium for the Cultivation of Trypanosomes. Z. Parasitenk. 41, 231–238 (1973). https://doi.org/10.1007/BF00329489
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DOI: https://doi.org/10.1007/BF00329489