[Studies on the vitality of human bone marrow during collection and deep-freeze preservation]. / Untersuchungen zur Vitalität von Human-Knochenmark bei der Gewinnung und Tieftemperaturkonservierung.

The cryoprotective effect of dimethylsulfoxide, glycerol and polyethylenglycol during freezing and thawing of human bone marrow was investigated by eosin staining test, an acridinorange fluochrome staining test and by RNA- and DNA-synthesis tests. In these tests the overall yield of vital nucleated cells, referred to the number in the absence of cryoprotectants and freezing and thawing, amounted to 50% with dimethylsulfoxide, 30% with glycerol, and 10% with polyethylenglycol. With dimethylsulfoxide and glycerol the loss of vital nucleated cells is almost entirely due to the addition of cryoprotectants. Polyethylenglycol freezing and thawing also leads to a great loss of vital nucleated cells. The results with dimethylsulfoxide show that the currently employed techniques of punction, preparation, freezing and thawing of bone marrow are suitable for clinical application.

[A test system for the measurement of cellular RNA-synthesis in the human bone marrow]. / Ein Testsystem zur Messung der zellulären RNS-Synthese von humanem Knochenmark.

A test for the cellular RNA-synthesis (incorporation of 3H-uridine in the RNA) of human bone marrow has been standardized with respect to the time of incorporation, the number of cells and the concentration of 3H-uridine. The following parameters were estimated for 500 microleter standard assay and 100 microleter aliquots for the determination of the radioactivity: time of incubation 80 min, number of nucleated cells 8 - 10(5), concentration of 3H-uridine 8,3 - 10(-6) M. Actinomycin D inhibits the RNA-synthesis to 90% in a concentration of 1.2 - 10(2) microgram/ml. The test appears generally applicable for the determination of the vitality of bone marrow after cryopreservation, the testing of cryoprotectants and haematotoxic substances and the control of the reaction of the bone marrow during chemical- or irradiation treatment of tumors.

[The effect of inosine, inorganic phosphates and pyruvate on erythrocyte glycolysis metabolites under conditions of preservation]. / Der Einfluss von Inosin, anorganischem Phosphat und Pyruvat auf die Glykolysematabolite in Erythrozyten unter Konservierungsbedingungen

Human erythrocytes were stored as resuspensions in solutions containing citrate (Z), inosine + citrate (I), inosine + phosphate (IP), and inosine + phosphate + pyruvate (IPP). The storage was made at + 4 degrees C for 6 weeks; the initial pH-value amounted to 7.4 at + 4 degrees C. The cellular concentrations of 2.3 DPG, ATP, G6P, FDP and DOAP + GAP were determined. The following results were obtained: 1. During the storage in stored Z-blood the 2.3 DPG concentration will fall below 10% of its initial value; it will remain nearly unchanged in stored I-blood and will increase to 170% in stored IP-blood, to 270% of its initial value in stored IPP-blood. 2. The ATP concentration of cells will fall to about 50% of its initial value at the beginning of the storage of all stored blood. After that it will only increase to about 80% of its initial value in stored IP- and IPP-blood. 3. During the storage the G6P concentration will increase to the highest degree in stored IPP-blood and if high pyruvate concentrations are not present, it will have a reciprocal behaviour towards the FDP and triosephosphate level. The results were discussed in view of the regulation of glycolysis under storage conditions.

[Autologous transfusion of erythrocytes with high or low concentration of 2,3-diphosphoglycerate. Survival rate and time and hemoglobin-oxygen affinity]. / Autologe Transfusion von Erythrozyten mit erhöhter bzw. erniedrigter 2,3-Diphosphoglyzerat-Konzentration. Uberlebensrate, -zeit und Hämoglobin-Sauerstoff-Affinität

1. In human erythrocytes the 2.3 DPG concentration was increased three to fourfold of the norm as IPP re-suspension by an incubation time of four hours at 37 degrees C or as ACD-AG blood was lowered below 20% of the norm respectively. After an autologous transfusion the 24 hours' surviving rate and the apparent half survival time of cells as well as the affinity of haemoglobin to oxygen in the total blood were measured. 2. The 24 hours' surviving rate for fresh erythrocytes with increased 2.3 DPG and ATP concentration amounts to 73% and the apparent half survival time amounts to 6 days. If erythrocytes are stored for four weeks as IPP resuspension at 4 degrees C, the 24 hours' surviving rate is 59%. Erythrocytes from fresh ACD-AG blood with lowered 2.3 DPG and a normal ATP concentration have a 24 hours' surviving time of 85% and an apparent half survival time of 24 days. 3. After autologous transfusion of 400 ml of erythrocytes with increased 2.3 DPG concentration the P50 value of the total blood will increase by 3 mm of Hg, after administering 400 ml of erythrocytes with lowered 2.3 DPG concentration it will fall by 1.8 mm of Hg. 4. The findings are discussed in connection with the significance of the changes of affinity of haemoglobin to oxygen produced by 2.3 DPG for the oxygen supply of tissues and under the aspect of using stored blood with increased 2.3 DPG concentration for practical purposes.