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A method for three-dimensional culture and biochemical stimulation to induce active platelet production

A technology of platelets and thrombopoietin, which is applied to animal cells, vertebrate cells, blood/immune system cells, etc., can solve problems such as difficult large-scale production, no obvious advantages in platelet activity detection, and cumbersome operation steps, etc. Strong operability and repeatability, promote the maturation of megakaryocytes, and improve the effect of production

Active Publication Date: 2018-03-09
血源生物科技(天津)有限公司
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Problems solved by technology

This 3D system is suitable for the growth of umbilical cord blood stem cells and directional induction into platelets, but the operation steps are relatively cumbersome, the culture period is relatively long, it is difficult to achieve large-scale production, and platelet activity detection (including platelet-like particle adhesion rate, CD62p expression Positive rate) has no obvious advantage
In addition, the system uses OPTI-MEM reduced serum medium, which cannot avoid the heterologous body in animal serum, and its platelet products may have the risk of human heterologous body rejection

Method used

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  • A method for three-dimensional culture and biochemical stimulation to induce active platelet production
  • A method for three-dimensional culture and biochemical stimulation to induce active platelet production
  • A method for three-dimensional culture and biochemical stimulation to induce active platelet production

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Embodiment Construction

[0038] The present invention will be further described below through specific examples, but the protection scope of the present invention will not be limited.

[0039] CD34 from umbilical cord blood + A method for cell differentiation to produce active platelets, the method comprising the steps of:

[0040] 1) Cord blood CD34 + Cell Separation

[0041] a. Pour the cord blood into a plasma bottle or a 50ml centrifuge tube, follow V 脐带血 :V 分选buffer =1:1 Add sorting buffer (recipe in Table 1), add 1 / 4 of the above mixing volume of hydroxyethyl starch and mix well, let stand at room temperature for 45 minutes;

[0042]

[0043] b. Transfer the supernatant to a 50ml centrifuge tube, centrifuge at room temperature and 1500rpm for 10min; discard the supernatant, add sorting buffer to resuspend the cells; take a 15ml centrifuge tube, add 4.5ml Ficoll separation solution to each tube (V Ficoll :V 分选buffer =1:1), slowly add the above-mentioned resuspension to the Ficoll separat...

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Abstract

The invention relates to a method for three-dimensional culture and biochemical stimulation to induce the production of active platelets. The invention utilizes a rotary three-dimensional bioreactor RCCS to co-culture CD34+ cells with staged small molecule compounds and cytokines, and can efficiently obtain active platelets ( CD41a+CD42b+). Compared with the prior art, the present invention has the following beneficial effects: the platelets obtained by the present invention have higher activity, stronger operability and repeatability, and can realize high-efficiency and rapid generation of functional platelets in vitro.

Description

technical field [0001] The invention relates to a method for inducing human umbilical cord blood CD34 + A method for cell differentiation to produce active platelets, specifically, the present invention relates to a three-dimensional culture and biochemical stimulation to induce human umbilical cord blood CD34 + A method for efficient cell differentiation to produce active platelets. Background technique [0002] Platelets are produced by megakaryocytes in bone marrow hematopoietic tissues and play an important role in the body's hemostasis and thrombus processes. The direct transfusion of platelets is an effective means to alleviate the low platelets in patients with blood diseases. However, domestic "blood shortage", blood product contamination, immune rejection and other problems lead to insufficient source of platelets, which brings troubles to the treatment of blood diseases. Therefore, seeking new sources of platelets and large-scale production of functional platelet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078
Inventor 周家喜杨亦青刘翠翠王洪涛苏培许元富姜尔烈
Owner 血源生物科技(天津)有限公司
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