CONSERVATION OF BLOOD (Latin conservare to store, keep) — methods of storage of blood out of an organism in a condition of its biological and functional full value. At conservation blood does not lose sterility, liquid properties during a certain term that allows to prepare and apply it to transfusion with to lay down. purpose.
The Russian scientist V. V. Sutugin for the first time in 1865 stated the idea To. to. for the purpose of its subsequent use at a military injury. In 1867
B. Rautenberg suggested to prevent a blood coagulation by means of addition of sodium carbonate. Systematic studying of problem K. to. in the USSR began in 1926 in Moscow, in first-ever Research in-those hematology and hemotransfusion. The big contribution to development of this problem was made D. N. Belenky, by C. I. Spasokukotsky, A. A. Bagdasarov, S. D. Balakhovsky, A. N. Filatov, S.E. Severin, F. R. Vinograd-Finkel, P. M. Maximov and many others.
In the 30th for To. to. began to apply TsIPK liquid — 5% solution of citrate to small cultivation (1:9) and the TsIPK glyukozotsitratny preservative No. 1.
By 1940 in the USSR the main objectives of problem K were resolved. to. also is established to lay down. efficiency of such blood that allowed to implement widely in practice its mass preparation. During 1941 — 1945 ways of prevention of bacterial pollution of blood at its mass preparation and storage were developed, and also the new preserving environments with citrate sodium citrate and antiseptic agents. It considerably increased quality To. to. also reduced danger of hemotransfusionic complications.
In the next years the theoretical and practical researches directed to lengthening of shelf-lifes of blood at temperatures continued it is above also lower than 0 ° — in the liquid and frozen state.
Conservation of blood at temperatures over 0 °
Conservation of blood at temperatures over 0 ° is carried out on purpose: 1) to stabilize in vitro blood in liquid state, i.e. to protect from coagulation, binding or destruction of any its component; 2) to keep funkts, activity and morfol. structure of blood cells, the exchange processes allowing to support on datum level or close to normal in them and circulation in circulatory system of an organism; 3) to keep ability to survival in an organism of the recipient after a transfusion; 4) to keep blood in a sterile state.
As stabilizers received the greatest distribution lemon to - that and citrate sodium citrate. The mechanism of their action consists in binding of calcium ions that prevents a blood coagulation.
Disodium salt of ethylene diamine tetraacetate — EDTANA 2 — also connects calcium ions. However at the same time it causes binding of potassium ions and magnesium and early hemolysis of stored blood that limited its use. Heparin (50 — 60 mg on 1 l of blood) is used for stabilization of blood of hl. obr. in cardiopulmonary bypasses. A shortcoming it is restriction of terms of stabilization (to 24 hours) and formation of clots at the expense of an inactivation of heparin in this connection it is applied only for short-term (several hours) To. to.
Stabilization of blood can be reached also without addition of chemical substances — by a transmission of blood through a column with cation-exchange resins. By this principle in the Belarusian scientific research institute of hemotransfusion E. D. Buglov in 1969 developed drug M - 1-phosphate of cellulose.
For To. to., except stabilization, preservation morfol, integrity of erythrocytes and them funkts, full value matters. For this purpose continuous inflow of the main substrate of food of these cells — glucose, and also means, obespechivashchy its utilization — enzymes and coenzymes is required.
The feedforward of oxygen and transport function of erythrocytes is established with contents 2,3 diphosphoglycerates (2,3-DFG), its important role in regulation of affinity of hemoglobin to oxygen and in the course of return of oxygen to fabrics: at low concentration in erythrocytes 2,3-DFG affinity of hemoglobin to oxygen is increased, at the same time dissociation of oxyhemoglobin and transfer of oxygen to fabrics are complicated; at high concentration 2.3-DFG communications of hemoglobin with oxygen are weakened, oxyhemoglobin dissociates quicker and fabrics easily remove oxygen from its complex with hemoglobin. It is known that ATP, except participation in formation 2,3-DFG, can be also connected with hemoglobin and influence process of return of oxygen to fabrics; therefore correlation of oxygen and transport function of erythrocytes with contents 2.3-DFG and ATP is supposed. Thus, along with stabilization, the main requirement to haemo preservatives — to fill up a lack of ATP and 2,3-DFG. Introduction to citrate solution for To. to. glucose gave the chance of extension of synthesis of ATP and a covering of need of erythrocytes for energy. Besides, an important point was finishing pH of the preserving solutions to 4,5 — 5,1 (at the same time erythrocytes consume glucose more slowly) that postpones approach of hemolysis and increases posttransfusion survival of erythrocytes.
Acid glyukozotsitratny solutions since 1947 gained recognition in many countries. They allow to keep stored blood at t 4 — 8 up to 21 days with posttransfusion survival of 70% of the poured erythrocytes that is the international standard. In the USSR TsOLIPK-76, LIPK-L-6 solution, in the USA, etc. the countries — ACD solution is applied.
Structure preserving solution and TsOLIPK-76: citrate sodium citrate — 2 g, glucose — 3 g, levomycetinum — 0,015 g, the bidistilled water to 100 ml. A shelf-life — up to 2 years.
Composition of LIPK-L-6 haemo preservative: citrate sodium citrate — 2,5 g, glucose — 3 g, sodium a sulfacetamide — 0,5 g, Trypaflavinum neutral — 0,025 g, the bidistilled water to 100 ml. A shelf-life — up to 7 days.
Acid glyukozotsitratny solutions became a basis for creation of new haemo preservatives, napr, the tsitroglyukofosfat containing 1 g lemon to - you, 0,75 g of a trinatriyfosfat, 3 g of glucose, to 100 ml about a distilled water, normal solution of caustic soda to pH 5,7 (20 ml of solution on 80 ml of blood) allowing to extend safety funk. full value of erythrocytes. Citrate and phosphatic solution with a dextrose — SRV is abroad applied.
Inclusion in haemo preservatives of metabolites of carbohydrate and phosphorus exchange (adenine, inosine, pyruvate, etc.) opened new perspective in To. to. — a possibility of recovery («rejuvenation») of tinned erythrocytes after marginal terms (21 days) of storage. An incubation it is long the stored tinned erythrocytes with metabolites of carbohydrate and phosphorus exchange leads to recovery lost in the course of storage them funkts, full value (contents 2,3-DFG, ATP, P50 and other indicators). The subsequent freezing of the recovered erythrocytes in liquid nitrogen allows to keep the «rejuvenated» cells a long time.
Methods of receiving and conservation of components of blood are developed: eritrotsitny, trombotsitny, leukocyte weight and plasma. Conservation of blood cells is of great importance, especially in connection with the increasing use in to lay down. to practice of transfusions of separate components instead of whole blood.
Eritrotsitnaya weight (see) — the most widespread Transfusion environment. Receive it by aseptic removal of plasma after upholding or centrifuging of stored blood; in the subsequent storage konts is possible. eritrotsitny weight (a hematocrit to 70%) - In to lay down. to practice also washed eritrotsitny weight (subjected to repeated aseptic washing fiziol is applied. solution), especially at reactive patients (sensibilized, allergized, etc.).
For use in a wedge, practice of leukocyte and trombotsitny masses methods of their receiving with use of centrifuging in the plastikatny equipment are developed and colloid besiege leu. Leukocyte weight (see. Leykokontsentrat ) hour remains to the 24th.
Trombotsitnaya weight (see) remains in own plasma at t ° 4 ° during 6 — 8 hours, and at t ° 22 ° in plastikatny bags — 72 hours.
Allocation and storage of leukocyte and trombotsitny masses, except the plastikatny equipment, carry out by means of special fractionators for automatic aseptic division of blood into components and receiving in large numbers of these cells from one donor by method of a cytapheresis. (see. Plasma exchange ).
Mass preparation of stored blood and its components is carried out by institutions of service of blood (the station and department of hemotransfusion) by uniform methodical rules. Blood is preserved by hl. obr. on the sterile haemo preservatives 76 and a tsitroglyukofosfata produced at the plants. Capture of blood from donors in glass bottles or plastikatny bags with haemo preservative is made in stationary operating rooms of stations and departments of hemotransfusion or in the place of work of donors where the special crews equipped with all necessary for preparation of blood go. Sterility To. to. it is provided with observance of strict measures of an asepsis at its preparation from donors, use of the sterilized haemo preservatives (in hermetically the corked bottles or plastikatny bags) and the closed sterile systems for capture of blood (fig. 1). Sterility and quality of stored blood strictly control by selective bacterial, the crops made at stations and in departments of hemotransfusion and also macroscopic assessment in to lay down. institutions before delivery for a transfusion.
Conservation of blood at temperatures below 0 °
Long-term storage of blood cells and plasma perhaps only at negative temperatures. At the same time cells remain in an anabiotic state — at suppression of metabolism, but preservation of activity of fermental systems and viability of cells.
Freezing and storage of plasma make at t ° — 30 °. The solution of the problem of freezing of erythrocytes was helped by establishment of the fact that ultrarapid cooling of blood (100 ° in a second) can occur almost without crystal hardening (in a thin coat it was possible to freeze and keep small volumes of erythrocytes), and discovery of cryoprotective property of the glycerin which allowed during the mixing with erythrocytes to keep them in the frozen state. A. D. Belyakov et al. in 1956 and F. R. Vinograd-Finkel with soavt, in 1958 developed a method of preservation of cellular elements of blood in the overcooled state at temperatures below 0 ° (from — 8 to — 16 °) without grain formation.
In practice apply two methods of cryoconservation of erythrocytes: ultrasnap-freezing in liquid nitrogen (— 196 °) with small (15%) concentration of glycerin or slow freezing with big (30 — 50%) concentration of glycerin at moderate temperatures (— 40 — 80 °) in an air chamber of electrorefrigerators. These methods allow is long to keep the unimpaired 85 — 95% of erythrocytes (for years).
The technique of ultrasnap-freezing of eritrotsitny weight consists that from whole donor blood after centrifuging erythrocytes allocate with observance of strict conditions of an asepsis and mix them with the sterile protecting solution containing glycerin. Mix is transferred to an aluminum corrugated container or a special plastikatny bag and subjected within 2 min. to freezing by immersion in a bathtub with liquid nitrogen then transfer to the special bunker or the camera also with liquid nitrogen for the subsequent long-term storage. For use of the frozen erythrocytes containers or bags take out from zhidkoazotny storage, subject to thawing by the placement on 25 sec. in a bathtub with water (t ° 45 °). After thawing make washing of erythrocytes from glycerin mannitno-salt, glyukozomannitny, salt and other solutions with consistently decreasing hyperosmia to an izoosmiya. For washing of erythrocytes use a method of consecutive centrifuging and draining nadstoya or automatic fractionators of various type for aseptic washing of the defrozen erythrocytes in a loop system. The washed erythrocytes fill in with the equal volume of isotonic solutions (sakharozoglyukozofosfatny, salt, etc.) then they are suitable for transfusion during the 24th hour.
The technique of slow freezing at moderate temperatures has the advantages — the zhidkoazotny equipment since electrorefrigerators are used is not required. As an endocellular kriofilaktik widely apply glycerin in big concentration (40%) though it and complicates ways of its washing after defrosting of erythrocytes.
For cryoconservation of leukocytes and thrombocytes the protecting solutions containing kriofilaktik endocellular (glycerin, a dimethyl sulfoxide, dimethylacetamide) or ekzotsellyulyarny action (polyvinylpirrolidone) in combination with carbohydrate solutions are picked up (sucrose, glucose, ascorbic to - that). Freezing of cells is made in specially designed devices allowing to cool them according to the set program. After program freezing and thawing of leukocytes it is possible to receive from 75 to 92% of the recovered cells.
There is a method of division of leukocyte weight into lymphocytes and granulocytes. The defrozen lymphocytes intend for typing and transfusion by the patient with oppressed immunol, activity; granulocytes can be used at treatment of patients with septitsemiya, an agranulocytosis, etc.
Freezing, storage, thawing and washing of erythrocytes and other blood cells are made in specially organized departments (banks) of long-term storage of the frozen blood at institutions of service of blood (fig. 2).
Klien, experience confirms efficiency of transfusions of a suspension of the defrozen erythrocytes at treatment of acute blood loss, anemias of various etiology, at open heart operations and during the use of the device «artificial kidney». Advantages of transfusions of the defrozen washed erythrocytes consist in their best portability (without posttransfusion reactions) the sick, sensibilized or allergized previous hemotransfusions or medicamentous means. They do not contain immunoaggressive cellular (leukocytes and thrombocytes) and the proteinaceous components of plasma which are the main reason for reactions at repeated transfusions (see. Hemotransfusion ).
The method of cryoconservation of blood, in addition to ensuring long-term storage, creation of reserves of blood of rare groups, reduces risk of infection with a viral hepatitis of Century. It also gives the chance of broad use of autotransfusions by preliminary accumulation of blood from this patient and its long-term storage in the frozen state until operation or need of transfusions (see. Autohemotrasfusion ).
the Idea of preparation and use posthumous (cadaveric, post-agonal, fibrinolizny) blood was stated and experimentally proved by B. N. Shamov in 1929 who proved that blood of corpses of animals in the first 6 — 8 hours after death keeps the full value has no toxic properties and renders to lay down. effect at transfusion to bloodless dogs. In 1930
C. S. Yudin for the first time with success made transfusion of posthumous blood to the patient with acute blood loss. Further long-term researches of a number of authors formed the basis for the conclusion about safety funkts, full value of posthumous blood, its non-toxicity and expressed to lay down. efficiency for the person.
Special quality of posthumous blood is its ability after preparation to be curtailed, and then «to be developed», i.e. the clot turns into liquid state again. This property called fibrinolysis (see) — difficult biol, the process happening in system of a blood coagulation after death. It is used with the diagnostic purpose: the fibrinolysis is characteristic only of blood of suddenly died people and is not observed in cases of death after a long agony. The fibrinolysis allows to prepare and store blood without addition of the stabilizing means. In the USSR in the Moscow city Scientific Research Institute of Emergency Medicine of N. B. Sklifosovsky and the Leningrad city Scientific Research Institute of Emergency Medicine of the prof. Yu. Yu. Dzhanelidze the wide experience of preparation and use of posthumous blood is accumulated. In a number of the cities special separations for preparation from corpses of bodies and fabrics, including blood are organized. There are certain rules of capture of blood (in the first 6 — 8 hours after death) from suddenly died as a result of acute cardiovascular insufficiency, a spasm or a sclerosis of coronary vessels, a myocardial infarction, an idiopathic hypertensia, a hematencephalon, an electric trauma, closed injury of a skull, a spinal cord, cervical department of a backbone, asphyxia from a prelum. The possibility of lengthening of shelf-lifes of posthumous blood by addition of the special preserving solutions (sakharozoglyukozofosfatny) is established.
Use of the posthumous blood stored at a temperature of 4 — 6 ° is allowed after obtaining results of a laboratory blood analysis and court. - medical necropsies. By data K. S. Simonyana et al. (1975), apprx. 24% of the prepared blood is rejected: 7% on bacterial pollution, 13% on serol, to indicators and about 4% on the basis of contraindications according to a pathoanatomical research.
Transfusion of posthumous blood was not widely adopted in to lay down. to practice of hl. obr. in connection with the good organization of donorship and service of the blood providing to lay down. institutions by stored blood.
See also Conservation of bodies and fabrics
Bibliography: Agranenko V. A. and Obshivalova of H. N. Metod of vosstanov-l of an ena I (ohm of l marry I) to an onser vir ov anny erythrocytes of deadlines of storage, Owls. medical, No. 8, page 66, 1976; Agranenko V. A., etc. The comparative characteristic of eritrotsitny weight and the whole blood prepared on different ge-mokonservant, Probl, gematol and a modulation. blood, t. 21, No. 8, page 37, 1976; In i-nograd-Finkel F. R., etc. Conservation of blood at temperatures below 0 °, in the same place, t. 3, No. 1, page 27, 1958; Gavrilov O.K. Scientific and organizational bases of service of blood, M., 1977; Golosova T. V., Kiselyov A. E. and M and r about l and N and A. N. Aseptik at preparation of stored blood, M., 1974, bibliogr.; The application guide of blood and blood substitutes, under the editorship of A. N. Filatov, L., 1973; With and m about-nyan K. S., at t and about N of t about in and K. P. and Tsurinov E. G. Posthumous blood in "aspect of transfusiology, M., 1975, bibliogr.; III and the m about in V. N. Problem of transfusion of cadaveric blood, Is new. hir. arkh., t. 36, book 3-4, page 581, 1936; Yudin S. S. Chosen works, book 2, page 309, M., 1960.
V. A. Agranenko.