Method for preparing PL (platelet lysate)

A platelet lysate, platelet technology, applied in the field of cell biology, can solve the problems of transplantation failure, immune rejection, lack of comparability, etc., to achieve the effect of increasing the scope, reducing waste, and improving the lysis rate

Pending Publication Date: 2017-11-24
重庆赛纳思生物科技有限公司
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  • Abstract
  • Description
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Problems solved by technology

However, FBS has its unavoidable disadvantages: (1) MSCs may introduce heterogeneous components during the culture process, resulting in immune rejection and resulting in transplantation failure; (2) zoonotic infectious diseases that may be introduced by FBS
The current research mainly adopts the preparation of isolated and concentrated platelets or apheresis from the peripheral blood of a small number of volunteers, which cannot be scaled up to meet the needs
Therefore, it also brings abou

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  • Method for preparing PL (platelet lysate)
  • Method for preparing PL (platelet lysate)
  • Method for preparing PL (platelet lysate)

Examples

Experimental program
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Embodiment

[0035] The method for preparing platelet lysate of the present invention comprises the following steps:

[0036] Step 1: Centrifuge the concentrated platelets in a centrifuge at 800-1000rpm for 15-25min. After centrifugation, the upper layer is platelet-rich plasma, and the bottom layer is red blood cell layer. The bottom red blood cell layer is separated. The concentrated platelets are derived from 400mL peripheral blood PC of voluntary blood donors, which are discarded after being shaken and stored at 22±2°C for 5 days, and then frozen for use. Utilize resources and reduce waste of resources.

[0037] Step 2: The platelet-rich plasma in the upper layer is fully mixed in batches, and the platelets are fully lysed by repeated freezing and thawing.

[0038] In the operation experiment, under PL freeze-thaw conditions, multiple PCs were evenly mixed, and were divided into 3 groups according to the procedures of -80°C freezer 8h / 12h / 24h and 37°C air bath thawing, each group was ...

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Abstract

The invention discloses a method for preparing a PL (platelet lysate) and an application of the PL. The method for preparing the PL comprises the following steps: concentrated platelets are centrifuged under a centrifuge at 800-1000 rpm for 15-25 min, the upper layer is platelet-rich plasma and the bottom layer is a red blood cell layer after centrifugation, and the red blood cell layer at the bottom is separated out; the upper platelet-rich plasma is mixed thoroughly in batches, and platelets are sufficiently lysed with a multigelation process; cell debris is removed, and the final PL is obtained. The invention also provides a cell culture method for culturing cells with the prepared PL. With adoption of the method, the yield of PL extracted from the concentrated platelets can be increased to 95%, operation time is shortened, and clinical expired products can be repeatedly used and developed.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a method for preparing platelet lysate. Background technique [0002] Mesenchymal stem cells (mesenchymal stromal cells, MSCs) have broad application prospects in cell therapy, tissue engineering, regenerative medicine, immunity and inflammation regulation. MSCs derived from bone marrow, umbilical cord blood, fat and dental pulp still need to be expanded to a certain number in vitro before clinical application. At present, fetal bovine serum (FBS) is the most commonly used supplement for in vitro culture of stem cells. As a heterogeneous source, FBS may bring disadvantages such as immune response and carrying heterogeneous pathogens, and its uniformity has also been widely questioned. Platelet α granules are rich in chemokines and growth factors, such as PDGF, TGF-β, IGF-1, SDF-1, VEGF, EGF, bFGF, BMP-2 / 4 / 6, etc., during the activation process through degranula...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2500/84
Inventor 邓小军
Owner 重庆赛纳思生物科技有限公司
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