DD152719A1 - METHOD OF PRESERVING ERYTHROCYTES - Google Patents

Abstract

The invention relates to a method for the preservation of human and animal erythrocytes. The object of the invention is to develop a facilitated manufacturing process for preservation. According to the invention, a glucose-sucrose-citrate-NaCl solution with or without the addition of adenine and optionally also guanosine, which may also contain other substrates and oxidizing agents, is added to the preservation solution.

Description

- ^Λ ~ 223 655

Dr. D. Strauss

Β _β pawl. _ · _

Eg Seidel.

"Method of preservation of erythrocytes ^

Field of application of the invention

The invention relates to a method for the preservation of human and animal erythrocytes, preferably of leucocyte and platelet-poor packed red blood cells for a period of up to 6 weeks at +1 ⁰ C to +6 ⁰ C. The method is applicable in the transfusion service. The result is clinically used for improved hemotherapy

Char akteristik the known technical solutions, the growing need for transfusions buffy poor packed red blood cells from human blood is primarily covered by non-conserved and therefore not lagerfäliige products that are made to 'different way-:

1 »One to several washings of erythrocyte concentrates stored for 1-7 days * The erythrocyte concentrates, which are subsequently resuspended in isotonic HaCl" solution with or without nutrient addition, are intended for immediate clinical use.

(Strauss, D .; Starck, E., Seidel, B ₀ J Zschr _o inn. Med.

submitted for publication) 2 * Erythrocyte concentrates filtered through cotton filters and suspended in isotonic IJaCl solution are also suitable for

immediate clinical use determined · (Prins, H, K _0; Henrichs, H "P" .J _e; Conemans, J, M _e H _e; Lenrink, _H6 J _9: 17th Congress ISH and 15th Congress ISBT. Paris ·

197 & * Kikugav / a, K .; Minoshima »K .: Vos Sang. 34 (1973), • 281 ~ 290) ""

Frozen preserved packed red cells are storable for a maximum of 24 hours after thawing, washing and resuspending, and are only available to a limited extent (Toursei, Wi; Dobslaw, B *; Scheibe, I _e ; Kaatz, H, Dt. Ge. Sundh -Wesen 33 (1978), 1142-1147)

4 * The by centrifugation and removal of the upper, predominantly leukocytes and platelets-containing cell layer (buffy coat) are prepared erythrocyte concentrates by resuspension in isotonic solution immediately UACL transfundier © n ₀

(Boskma, R _e; Huges, R .; McShine, R * L _#, Smit Sibinga C ₀ T ,; 17th Congress ISH and 15th Congress ISBT, Paris 1978 "Prins et al ibid _e)

When suspended in autologous plasma, which in this case acts as a preservative, eine'mehrwöchige storage (Smit Sibinga, CT *, Bosnia, J _e; 17th Congr * ISH and 15 th Congr · _e ISBT Paris, 1978) is possible

However, this method results in the loss of plasma, which is otherwise urgently needed for the production of valuable drugs and is not required for the preservation of erythrocytes _e

5 »In the GDR - as the first buffy coat in the world - poor RBC concentrates are routinely produced and after resuspending in a sugar-electrolyte solution (CDS-AG solution) according to the provisional quality regulation (IPAR) up to 6 weeks storage ψ 4 ⁰ C clinically used »

(Strauss, D *; Seidel, B. Meurer, W. Markova, ΝοΑ _β : Folia Haematol.107 (1980), 104-124) Disadvantages of this procedure are:

- High sucrose concentration in the solution, 234 mmol / 1, which causes technical difficulties in the preparation of a non-caramelized solution, free of insoluble impurities and, moreover, an additional burden on the patient with fiber ».

223 655

Absence of inorganic phosphate in the ODS-AG solution, which, however, is required for the preservation of the erythrocyte t.off change ₀ . ,

- High adenine concentration, which can be reduced by half to the same extent, if the vitality of the preserved cells is preserved «

(Strauss, 'D., Seidel, B ": Dt ₀ Gesundh.-Wesen 35 (1980), 28 - 3D ..."

- The one from Hb'gman / Sweden

Högman, CF .; Hedlund, K «; Zetterström, H »: Έ, Engl ₀ J ,: Med. 299 (1978), 1377-1382 .

Höginan _? CF; Hedlund, K .; Akerblom, 0 .; Venge, P; Transfusion 18 (1978), 233-241)

developed solution for resuspension of buffy coat-poor packed red cells (SAG) can be several weeks of storage at _o The disadvantage of this method is, however, that 'performs this consisting only of lacI, adenine, and glucose preservation solution to a higher haemolysis of the erythrocytes, - after 5 - 6 weeks Storage 3 - 4 % "This high rate of hemolysis is not acceptable for clinical use. A high rate of hemolysis. occurs in the composition of preservatives developed by Australian authors in the same composition as in Högman ", (Lovric, VA _e ; Prince, B ₀ , Bryant, J, Vox Sang 33 (1977), 346-352).

Ze d l he invention

- IMIM HIIIiWi tarn ι ii iiniil MaMHEiMCmn wn ywa ^

The object of the invention consists in a higher therapeutic effect of erythrocyte substitutions in the patient as well as in a simplified technical production process for the preservation solution and at the same time cost savings of 50 % of chemicals for the preservation solution. The object of the invention is the composition of the conservation solution routinely used in the GDR

for human erythrocyte concentrate (CDSrAG solution) in 6 positions in order to achieve the desired goal «

According to the invention, human erythrocyte concentrates are prepared from 1 "transfusion unit (400 ml blood + the respective preservation solution) ACD, ACD AG, EDTA, CPD, CPD-A, CPD-AG or citrated blood in VA to ^1/2 of the cell volume suspended with the new preservation solution "the new preservation solution contains with respect to the known solution, in addition Kacl and Uatriumphosphat and lesser amounts of sucrose (only VA) f optionally Adeninsulf at (only ^1/2). in addition, , may also contain other substrates and oxidizing agents. The sucrose may be replaced by mannitol or other disaccharides or sugar alcohols in quantities of 10-100 mmol / l and citrate alone in amounts of 1-6 mmol / l. It has been found that maintain the vitality of the red cells to the same extent as in previous stays preservation methods ₀ the low hemolysis rate of about "1% after 6 weeks of storage was by partial Austaus ch secured sucrose against UACL v / earth · The oxygen transport function of erythrocytes is by the addition of phosphate in the erythrocyte concentrate (2 mmol / 1), especially in red blood cell concentrate s of CPD, EDSA- or ₀ citrated long obtain "An initial waste des'extrazellulären pH _f which makes up 0.3 pH units in the high-sucrose medium is prevented «

The production of the preservation solution is facilitated by the strong reduction of the sucrose concentration to only VA in favor of HaCl addition and the caramelization of the solution occurring under routine conditions in the transfusion service during sterilization by means of autoclaving is substantially reduced.

Reduce the cost of the substances to be employed in order 50% ₉ are saved per 1 solution 60 g of sucrose and 0.25 g Adenineulfat _s at the present Produktionatim-

or more in the expected increase in production to 8 times the previous volume for this product «. , ,

A storage period of 3 weeks is possible with adenine-guanosine-free solution, with adenine 5 weeks and with further addition of guanosine 6 weeks at + 1 to + 6 ° C · Higher temperatures shorten the storage times. ¹ . ^

embodiment

400 ml of blood from a healthy donor are placed in a sterile 500 ml glass jar (or in a blood plastic bag) containing 50 ml of stabilizer consisting of Na ^ citrate 117, citric acid 5 and HaCl 259 mmol / l The bottle is centrifuged for 20 minutes at 700 g within a few hours after taking the blood. Plasma and then buffy coat are squeezed off and added to the remaining approximately 150 ml cell sediment in 100 ml preservative solution. The cells are resuspended by swirling the bottle several times _c

The starting B-pH value is 7.2, the pH value is 7.15 * The hematocrit is 0.6. After 6 days of storage, erythrocytes have an ATP content of 50 % of normal and in vivo survival rates of 70-80 % .

Claims (2)

2ΰ 6 5 5 Claim for invention ·:: Method of preserving you living human and animal erythrocytes with or without leucocytes and platelets, characterized in that a Gluko ™ se sucrose citrate-UaCl solution with or without the addition of adenine and optionally also guanosine, in addition also contains additional substrates and oxidants, added 2 «method according to item 1, characterized in that the sucrose in amounts of 10 to 100 mmol / 1 erythrocyte concentrate is used vdrd ₆ ' 3β Process according to item 1 and 2, characterized in that, instead of sucrose, wholly or partly mannitol or other disaccharides or sugar alcohols are used « 4 * Process according to items 1 to 3 », characterized in that, instead of sucrose or mannitol, the ITatrium citrate is used in amounts of 1 to 6 mmol / 1 of packed red cell concentrate« Method according to items 1 to 4, characterized in that sucrose or mannitol is also used together with citrate in the stated amounts _e 6 _e method according to item 1 to 5 $, characterized in that • as substrates for the erythrocyte metabolism xylitol, ribose or dihydroxyacetone Ό. Ä. added to maintain the 2,3-Bisphosphoglyzeratgehalts « 7. The method according to item 1-6, characterized in that a hydrogen donor for the hydrogenation of hikotin • a reamid-adenine dinucleotide (HAD) as z »B * pyruvate, ascorbate or methylene blue is used» 8. "method" as claimed in point 1-7, that the preservative solution is added in a volume of V4 to ^1/2 * cell volume " 9 "method according to item 1 ~ 8, characterized in that with adenine and guanosine-free preservative solution a storage period of 3 weeks and with adenine of 5 weeks ^ with simultaneous adenine and Guanosinzusatz of 6 Y / ochen at ψ 1 ° to + 6 ⁰ C is possible _for the storage periods at higher storage temperatures but reduce "