Studies In Blood Preservation

Transcription

1 Howard Unversty Dgtal Howard Unversty Faculty Reprnts Studes In Blood Preservaton Charles R. Drew Follow ths and addtonal works at: Part of the Medcne and Health Scences Commons Recommended Ctaton Drew, Charles R., "Studes In Blood Preservaton" (1939). Faculty Reprnts. Paper Ths Artcle s brought to you for free and open access by Dgtal Howard Unversty. It has been accepted for ncluson n Faculty Reprnts by an authorzed admnstrator of Dgtal Howard Unversty. For more nformaton, please contact lopez.matthews@howard.edu.

2 STUDIES IN BLOOD PRESERVATION 1 F a t e o f C e l l u l a r E l e m e n t s n R e l a t o n to P o t a s s u m D f f u s o n C h a r l e s R. D r e w, M.D., K a t h a r n e E d s a l l, a n d J o h n S c u d d e r, M.D., N e w Y ork, N. Y. IN A PREVIOUS publcaton1 we reported a tenfold rse n plasma potassum of preserved blood. Ths was not due to bacteral contamnaton. It was ncreased by trauma, such as shakng, and was defntely modfed by the area of the nterface between the sedmented cells and the overlyng plasma. The queston was rased as to whether ths ncrease n plasma potassum was a pure dffuson process or whether t was due to actual cell destructon. Ths aspect of the problem s reported here. m e t h o d Fve cubc centmeters of freely flowng venous blood were collected n each of 35 sterle round-bottomed test tubes contanng 5.0 mg. of heparn as an antcoagulant. After nvertng each tube three tmes, they were plugged wth cotton and kept n a refrgerator at approxmately 4 C. throughout the perod of the experment. Each day one tube was taken from the refrgerator, and centrfuged for one hour. After 0.5 c.c. of plasma had been removed for potassum analyss,2 and after the blood had been thoroughly mxed by nvertng the tube ffteen tmes, the followng determnatons were made: (1) Red cell count. (2) Hemoglobn (Hellge). (3) Whte cell count. (4) Dfferental whte cell count. (5) Platelet count. (6) Mean cell dameter of red blood cells (Halometer method). * *From the Surgcal Pathology Laboratory of the College of Physcans and Surgeons, Columba Unversty, and the Departments of Surgery Medcne, Presbyteran Hosptal, New York. ; Ths study was made possble by a grant from the Blood Transfuson Betterment Assocaton, New York. Receved for publcaton, Aprl 1, 1939.

3 d RESULTS The donor, A. J., was a professonal man of Internatonal Group 0. V enous B lood AT START OF AT END OF PHLEBOT- PHLEBOTOMY OMY (590 c.c.) Hematocrt per cent cells Plasma specfc gravty Plasma protens gm. per cent Whole blood potassum mg. per cent Plasma potassum 21.5 mg. per cent Cell potassum (calculated) mg. per cent ph 7.51 V alues on I ntal 5.0 c.c. Sam ple* Red blood cell count 5,500,000.0 Hemoglobn 14.2 gm. per cent Whte blood count 7,000.0 Dfferental whte cell count Polymorphonuclear leucocytes 52.0 per cent Eosnophlc leucocytes 2.0 per cent Basophlc leucocytes 1.0 per cent Lymphocytes 42.0 per cent Monocytes 3.0 per cent Platelets 140,000.0 Mean cell dameter 7.4 Plasma potassum 20.6 mg. per cent The results are graphcally represented n Charts 1 to 4. * Corrected for 0.5 c.c. plasma removed. DISCUSSION Red Blood Cells. The mantenance of the red blood cell counts at approxmately a constant level over a perod of a month suggests that, n the steady ncrease of the potassum content of the plasma, cell destructon plays at most only a small part. Duran Jorda and hs co-workers3 reported there was a loss of 1,500,000 erythrocytes over a perod of sxteen to twenty days n blood conserved n ctrate. They suggested the stroma of these destroyed cells mght play a part n some of the slght posttransfuson reactons. We have not found changes as great as they report. The fact that all of our tubes were centrfuged for an hour before each count may play some part n ths dscrepancy. Yudn,4 from Russa, has reported that there s no loss n ablty of cells so stored to carry oxygen, hence the chef bologcal functon of the red blood cells seems unmpared. Hemoglobn. The hemoglobn content remans constant. Ths confrms the fndngs of prevous wrters. The work of Amberson5 has ndcated that the part of the hemoglobn whch escapes from the cells to gve the usual pcture of hemolyss s stll capable of functonng normally. Volume Index and Dameter of Red Blood Cells. The volume ndex of the cells decreases n successve specmens. There s a dmnuton n whte blood cells, but the greater part of the decrease seems due to the dmnuton of the sze of the red blood cells. At the end of thrty days these cells may lose as much as 25 per cent of ther chef base, 20 per cent of ther hemoglobn, and decrease 20 per cent n dameter. Ponder0 has shown that red blood cells do not

4 RATE OF POTASSIUM DIFFUSION IN HEPARINIZED BLOOD Chart 1. Potassum dffuson n heparnzed blood. Each pont on the chart represents a determnaton done on dfferent test tube samples each day and s calculated to represent the actual amount of potassum n mllgrams n the plasma of 100 c.c. of blood. The values are approxmately the same as n prevous experments where daly determnatons were done on the same blood wth the fnal curve representng- a summaton of the ncrements. C E LLU LA R CHANGES IN HEPARINIZED BLOOD Chart 2. Upper thrd. Red cell counts vared between 6,200,000 and 4,800,000, the mean beng- 5,500,000. No actual loss n number of red blood cells. Mddle thrd. Hemoglobn values vared between 14.2 and 17.6 gm. per cent. The curve, allowng for slght concentraton due to evaporaton, represents a constant hemoglobn content. The drop after the ffteenth day may be accounted for by the gradually ncreasng amounts of hemogrobn removed n the plasma taken for potassum determnatons. Lower thrd. There was a gradual decrease n the sze of the red blood cells from an ntal 7.6 mcrons to a fnal 5.8 mcrons as the cells lost both hemoglobn and potassum. 5

5 always act as perfect osmometers, a fact whch he attrbutes to the possble loss of ther salt content. Whte Blood Cells. The total whte blood cell count rapdly dmnshes wth the polymorphonuclear leucocytes dsntegratng; most rapdly. The reported loss of half of the complement of the blood by the sxth or seventh day, and complete destructon n ffteen to twenty days, may be assocated wth ths loss of leucocytes.3 The lymphocytes are more resstant. At the end of thrty days they are easly recognzable when found, but there are so few that counts are uncertan. Ths holds true, lkewse, for monocytes. PER C.MM CELLULAR CHANGES IN HEPARINIZED BLOOD n r r r 1.r r L - f T O T A L WHITE COUNT L \ 1 A L \1 1 T A ] ~ j _ r 1 *' LfJ 1 n nv nv ,000 1,000 L \ N F U T R O P H II F.S - v \ I \ H - - F «> _J \ r r r r <V- LYM PHOCYTES \ x & MONOCYTES 1 - > r r J O DAYS Chart 3. Upper thrd. The total whte blood cell count fell 50 per cent n twenty-four hours, the cells lost shape and gradually dsappeared by the sxteenth d a y ; those remanng appearng only as gelatnous-lke clumps. Range 7,000 to 675 cells. Mddle thrd. The polymorphonuclear leucocytes showed earlest and most rapd changes ; ther nucle soon losng shape and dsntegratng. Range, 3,600 to 14 cells. Lower thrd. Lymphocytes and monocytes dsappeared at a slower rate and retaned ther shape, sze, and stanng better than the leucocytes, beng qute dstnct when found on the thrteth day. Basophlc and eosnophlc cells, whle too few to be charted, retaned ther shape, sze, and stanng qualtes. The latter were least changed and very dstnct on the thrteth day. Range, 3,100 to 654 cells. The eosnophles appear most resstant. The nucleus breaks up late and the granules reman unusually well defned. Duran Jorda3 has reported smlar fndngs. Thrombocytes. The platelets n the ntal venous blood sample numbered 140,000. These rapdly dmnshed n the frst three days and remaned at about -

6 1. In heparnzed preserved blood there s lttle or no actual loss n the number of red blood cells over a perod of thrty days. 2. The hemoglobn content remans constant n the total sample, though 15 to 25 per cent may be found n the plasma. 3. The mean cell dameter of the red blood cells s reduced approxmately 20 per cent n thrty days. 4. The polymorphonuclear leucocytes are dmnshed by 50 per cent n forty-eght hours and are amorphous masses n ffteen days. 5. The lymphocytes, monocytes, and eosnophles do not dsntegrate so rapdly, the latter are partcularly well preserved. 6. The thrombocytes rapdly fall to a low level, then reman constant at about 30,000 for about ffteen days. 7. Potassum n the plasma of heparnzed blood reaches a level ten tmes normal n thrty days. 7 30,000 for ffteen days. Centrfugaton may have some part n these counts. Ths suggests, however, that syndromes presentng thrombocytopena would not be benefted by blood whch had been stored too long. Potassum. The rate of potassum loss from the cells n the ndvdual test tubes s of the same order as that found n the flasks from whch daly samples were taken. The potassum content of whte blood cells (those obtaned from the buffy coat n case of lymphatc leucema) s greater than the red blood cells. In normal blood ther relatve number s nsgnfcant. However, ther early rapd destructon may account n part for the steep slope of the curve durng the frst few days. THROMBOCYTES IN HEPARINIZED BLOOD Chart 4. Thrombocytes rapdly fell to about 40,000 and for thrty days remaned approxmately at ths fgure. Only the frst three days are charted. CONCLUSIONS

7 REFERENCES 1. Scudder, J., Drew, C. R., Corcoran, D. R., and Bull, D. C.: Studes n Blood Preservaton, J. A. M. A. In press. 2. Truszkowsk, R., and Zwemer, R. L.: Cortcoadrenal Insuffcency and Potassum Metabolsm, Boclem. J. 30: 1345, Determnaton of Blood Potassum, Boclem. J. 31: 229, Duran Jorda, F., Aleu, E. M., Sarda Roca, J., Benlloch Llorach, A., Soteres Dez, F., and others: The Study of the Metabolsm of Ctrated Blood Conserved n Vtro, Surg., Gynec. & Obst., Internat. Abstr. Surg. 67: 386, Yudn, S. S.: Transfuson of Stored Cadaver Blood, Lancet 2: 361, Amberson, W. R.: Blood Substtutes, Bol. Rev. Cambrdge Phl. Soc. 12: 48, Ponder, E., and Robnson, E. J.: The Measurement of Red Cell Volume. V. The Behavor of Cells From Oxalated and From Defbrnated Blood n Hypotonc Plasma and Salne, J. Physol. 83: 34,