It may be concluded that the degree of dilution of the culture medium has a marked influence on the rate of growth of splenic tissue. The maximum acceleration was obtained in a medium composed of three volumes of normal plasma and two volumes of distilled water. The growth in this hypotonic plasma was very much larger than in normal plasma. On the contrary, the growth of the spleen in hypertonic plasma was always less than in normal plasma.
In other experiments, we found that in diluted plasma there was also an acceleration of the growth of the skin, the heart, and the liver of chickens. The skin of adult frogs also grew more actively in this plasma.
The optimum degree of dilution varied according to the nature of the tissues and to the species of the animals. While the plasma containing two fifths distilled water produced the largest growth of splenic tissue, a slightly less diluted medium was more favorable for the liver and the heart, and generally for the skin also. The action of hypertonic plasma varied also in a large measure. While the spleen did not grow at all in the medium containing 0.0124 and 0.0144 sodium chlorid, the skin, on the other hand, could stand a high concentration of the sodium chlorid. Even its growth was activated in media containing 0.0094 and 0.0124 sodium chlorid and was greater than with normal plasma. The spleen of kittens was very easily affected by the changes of the dilution of the plasma, while the skin of the frog presented its best growth in plasma containing one half distilled water. Marked variations in the sensitiveness of tissues to hypertonic and hypotonic media will probably be observed in animals of different species.
From these experiments, three conclusions can be drawn: namely, that certain laws of growth, discovered by Loeb, in lower organisms are true also for higher organisms; that normal plasma is not the optimum medium for the growth of tissue; and that each tissue has probably its optimum medium.
The growth of the spleen is, without doubt, considerably modified by the variations of the dilution and perhaps of the osmotic tension of the plasma. It is possible then that the influence of osmotic tension, discovered by Loeb, in the growth of certain organisms, is a general law applicable as well to higher forms of life— frogs, cats, and chickens—as to lower organisms—tubularia and sea-urchins. In placing tubularia in different dilutions of sea-water and distilled water, Loeb found that the greatest rate of regeneration was observed when two volumes of distilled water were added to three volumes of sea-water. But fertilized eggs of sea-urchins were more sensitive to the action of hypertonic plasma, and they all died in a dilution of sea-water with two fifths distilled water. If only one fifth distilled water was added to the sea-water they developed normally. We found that the cells of certain tissues of the chicken follow a similar rule, since the maximal growth of the spleen is obtained in plasma containing two fifths distilled water, while other tissues grow better in a less hypotonic medium.
Normal plasma is certainly not the ideal medium for the growth of tissues, since slight modifications of the tension, the alkalinity, or the addition of certain inorganic salts to normal plasma, increase the rate of the growth of tissues.