Small molecular sugar stable buffer used for blood platelet frozen-dried preservation

A technology of small molecule sugar and buffer solution, which is applied in blood diseases, freeze-drying transportation, preservation of human or animal bodies, etc., can solve the problem of low aggregation activity and achieve the effect of complete structure and normal shape

Inactive Publication Date: 2010-06-02
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, Brry SJ. et al. used glucose to incubate platelets and then freeze-dried and rehydrated, and obtained a cell recovery rate of 75%-85%, but the aggregation activity was low.

Method used

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  • Small molecular sugar stable buffer used for blood platelet frozen-dried preservation
  • Small molecular sugar stable buffer used for blood platelet frozen-dried preservation
  • Small molecular sugar stable buffer used for blood platelet frozen-dried preservation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0041] Trehalose was added to the basic buffer GS formula to prepare the load solution of GS (T35), which was a stable aqueous buffer solution containing 35mM trehalose, and its osmotic pressure was 339±8mOMS. The aqueous solution contains: trehalose 35mM, glucose 10mM, sucrose 12.5mM, sodium citrate 12mM, KCL 5mM, NaCL 140mM, PGE 1μg / mL.

[0042] According to the aforementioned 2) small molecule sugar loading experiment and 3) detection of various indicators of platelets loaded with small molecule sugar, the results:

[0043] ① Cell recovery rate: greater than 70%;

[0044] ② Aggregation activity: greater than 60%;

[0045] ③Morphological structure: The morphological structure is basically normal, see Figure 2b .

[0046] According to the aforementioned 4) the platelets were lyophilized and hydrated and the platelet samples obtained after 5) were freeze-dried and hydrated for physiological index detection, the results:

[0047] ① Cell recovery rate: greater than 70%;

...

experiment example

[0054] According to the recipe in Table 1, trehalose was added to the basic buffer to prepare loading solutions such as stable buffers GS (T30), GS (T40), and GS (T45). For example: GS (T40) stabilization buffer contains 40mM trehalose, and its osmotic pressure is 386±6mOMS.

[0055] At the same time, GS was used as a comparison sample in the experiment. Another trehalose solution (10 mM KCl, 10 mM EGTA, 10 mM imidazole, 1 μM PGE1, 100 mM NaCl, 40 mM trehalose) prepared by Wolkers WF. et al. was used as a control sample.

[0056] Table 1: Stabilization buffers containing different concentrations of trehalose

[0057]

[0058] Detect every index of the platelet of processing with the method identical with embodiment, result:

[0059] 1) Detection of various indicators of platelets loaded with small molecule sugar:

[0060] The results of ①cell recovery rate and ②aggregation activity are shown in Table 2. It can be seen that the recovery rate of platelets after treatment w...

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Abstract

The invention provides a stabilizing and buffering liquid of low molecular weight sugars used for lyophilization preservation of platelet, which is water solution with 35-45mM thickness by adding thelow molecular weight sugars on the basis of basic buffering liquid composing prescriptions (GS), wherein the basic buffering liquid composing prescriptions (GS) comprises 140mMNaCL, 5Mmkcl,12mM sodiumcitrate, 10mM glucose, 12.5mM sucrose and 1mug / mLPGE1. The low molecular weight sugars are trehalose. After the platelet is stabilized and protected by the stabilizing and buffering liquid of low molecular weight sugars of the invention, the platelet is freeze-dried with rehydration. The recovery ratio of a cell is above 70% and accumulating activity evoked by ADP can achieve 50% of fresh platelet. The platelet freeze-dried with rehydration has normal shape and complete structure, thereby a new solution is provided for freeze-drying conservation of the platelet.

Description

technical field [0001] The invention belongs to the field of freeze-drying preservation of platelets, and relates to a small molecule sugar stabilizing buffer. Background technique [0002] The study of freeze-dried platelet preservation is of great significance. Firstly, freeze-dried platelets have broad application prospects in treating the wounded and sick in hospitals. The demand for platelets in my country's direct market value is estimated to be about 1 billion per year (50-700,000 bags × 1700 Yuan), the indirect benefits cannot be estimated. And it is suitable for storage and mutual adjustment in various large, medium and small hospitals in different regions and remote island areas. Secondly, freeze-dried platelets have important military value. Soldiers can carry them with them for field rescue, save the lives of the wounded, and play a role in maintaining the combat effectiveness of troops. Thirdly, it can effectively prevent the spread of transfusion-transmitted d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078A61K47/26A61K35/14A61K9/19A61P7/00G01N33/53G01N33/50A01N1/02A61K35/19
Inventor 王捷熙韩颖杨超王艳刘敏霞权国波刘安
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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