Okayama Igakkai Zasshi (Journal of Okayama Medical Association)

Studies on the Tissue Culture of Rabbit's Bone Marrow

Part I. The Influence of osmotic Tension of Culture Media

Makoto Tsushima

1956 Volume 68 Issue 8special Pages 1-13
Published: August 31, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.8special_1

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The influences of osmotic tension upon the tissue culture of rabbit's bone marrow were investigated.
The proliferation of cultivated tissue was expressed and examined numerically by the proliferation-index which was introduced by the density and the proliferation area.
The cell proliferation was accelerated in the hypotonic medium but not so markedly as described hitherto. In the hypertonic medium, it was suppressed.
When the serum plasma was used as the culture medium in various dilutions prepared with Locke's solution, maximal proliferation of the tissue was observed at one fourth of the initial serum concentration.
In the hypotonic medium, the migration of pseudo-eosinophilic leukocyte looked somewhat irritated at first but soon it became tardy. On the other hand, in the hypertonic medium, the migration was suppressed a little at first but it gradually recovered its initial activity.
These facts seem to show that the physico-chemical direct effects of pharmaca should not be over-looked when the effects of pharmaca upon the tissue culture of rabbit's bone marrow is studied.

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Studies on the Tissue Culture of Rabbit's Bone Marrow

Part II. The Influence of Temperature

Makoto Tsushima

1956 Volume 68 Issue 8special Pages 15-25
Published: August 31, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.8special_15

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The influences of temperature upon the tissue culture of rabbit's bone marrow were investigated.
The effects of various temperatures from 3° to 37°C were tested by Carrel's bottle method, in which a specially designed apparatus was used in this experiment. The lowest temperature was 12°C at which the proliferation of cultivated tissue was scarcely ascertained. The proliferation was nearly proportional to the temperature-rise in the range of from 12° to 37°C.
Another test was carried out by the slide culture method from 33°C to 41°C. At 41°C proliferation was not observed any more, and the maximal proliferation was obtained at 39°C. But this proliferation at 39°C was not the normal one but somewhat irritated or abnormal one. Thus, for the ordinary observation, cultivation at 37°C is thought to be more suitable.
The migration rate of pseudo-eosinophilic leukocyte showed an increase in proportion to the ascent of temperature. But the great migration rates at high temperatures did not last long and fell very soon. The maximal migration rate was observed at 43°C. The increased migration rate lasted for at least 6 hours at these high temperatures. Up to 55°C the leukocytes did not lose their migratory power.
The cultured tissue could be kept alive for 16 days at 3°C but the proliferating power of the tissue decreased day by day.

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Studies on the Tissue Culture of Rabbit's Bone Marrow

Part III. The Influence of Hydrogen Ion Concentration of Culture Media

Makoto Tsushima

1956 Volume 68 Issue 8special Pages 27-37
Published: August 31, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.8special_27

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The influences of hydrogen ion concentration of media upon the tissue culture of rabbit's bone marrow were investigated.
On the course of culture the acidity of media gradually increased and in Carrel's bottle method pH-value changed from 7.80 to 7.06 in 6 days' culture.
When cultured in acid medium more prosperous proliferation was obtained with the use of comparatively lower acidity. Likewise, lower alkalinity proved more favorable for good proliferation.
In cases where good proliferations were obtained, the migration of pseudo-eosinophilic leukocyte was also vigorous.
Tyrode's solutions of various pH-values were prepared by adding adequate doses of sodium bicarbonate, and were used by mixing them with the media to find out the optimum pH-value for the tissue culture of rabbit's bone marrow. The maximal proliferation was obtained at pH 7.63 which was prepared by mixing Tyrode's solution of pH 8.00.
No significant changes in the migration rates of pseudo-eosinophilic leukocytes were caused by the added Tyrode's solution of various pH-values.
Thus the culture medium most favorable for the growth of tissue was decided as such of the hydrogen ion concentration of pH 7.63.

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The Influence of Cancer Tissue Extract on the Pancreas of Experimental Animals

Part I. The Influence of the Cancer Tissue Extract on the External Pancreatic Secretion

Hirosuke OMORI

1956 Volume 68 Issue 8special Pages 39-56
Published: August 31, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.8special_39

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The cancer tissue extract was made from surgically removed stomach cancer. A pancreatic fistula was made on dog and the change of the content of the various ferments in the pancreatic juice was studied after daily intravenous administration of 10% cancer tissue extract 1 cc/kg.
The following results were obtained;
1) Diastase was markedly decreased. The decrease began on the 1st day of the experiment, and the lowest value occurred in 2nd or 3rd day. This lowest value was maintained as long as the injection of the cancer tissue extract were continued. However, when injection of cancer tissue extract was discontinued, diastase values were gradually increased.
2) Trypsin also began to decrease on the 1st or 2nd day after injection of the cancer tissue extract. The lowest value occurred on the 2nd or 3rd day of the experiment. However, compared to diastase, this decrease was not marked.
3) There was found no marked change regarding the content of lipase.
4) In the control group, in which the extract of normal stomach tissue was used as injection material, there were found no change in the ferments of pancreatic juice.
The nucleic acid in the pancreatic tissue was studied histochemically by the Pironin-Methylgreen stain using rabbits which were given the cancer tissue extract 1 cc/kg daily intravenously.
The following results were obtained;
1) The amount of RNA was decreased moderately after daily injection of the cancer tissue extract for 2 weeks and marked decrease was found after daily injection for 4 weeks.
2) No significant changes were observed in DNA and KES.
It is believed that the function of external pancreatic secretion is definitely inhibited by the injection of the cancer tissue extract.

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The Influence of Cancer Tissue Extract on the Pancreas of Experimental Animals

Part II. The Influence of Cancer Tissue Extract on the Internal Pancreatic Secretion

Hirosuke OMORI

1956 Volume 68 Issue 8special Pages 57-71
Published: August 31, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.8special_57

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Insulin was extracted by the Sulfric acid-Alcohol method (Kumagaya) from the pancreatic tissue of dogs which were given 10% cancer tissue extract 1 cc/kg daily intravenously for various periods.
The amount of the extracted insulin was measured by the Toronto method using rabbits.
The following results were obtained;
1) The amount of insulin in the pancreas obviously decreased by the injection of cancer tissue extract compared to the control group.
2) Insulin began to decrease 1 week after the daily intravenous injection of cancer extract. The decrease was moderate after 2 weeks and marked after 4 weeks.
Pancreatic tissue of rabbits, which were given 10% cancerous extract 1 cc/kg daily intravenously, was studied histologically by Chromium Hematoxylin-Phroxin stain.
The following results were obtained;
1) It was found that the number of islets decreased; the size of islet became smaller; and the number of β-cells diminished.
Degenerative changes were found in the islets and β-cells after the daily intravenous injection of cancer tissue extract.
2) These changes were noticeable 1 week after the daily injection; moderate after 2 weeks; and marked after 4 weeks.
It is believed that the function of the internal secretion of the pancreas is inhibited by injection of cancer tissue extract.

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