Method for preparing PL (platelet lysate)

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...

AI Technical Summary

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 about the problem of PL uniformity

(2) So far, there have been nearly 200 clinical studies related to PL or PRP. However, due to the lack of strict standards for PL preparation and products, the preparation methods are inconsistent, resulting in huge differences between research results, lack of comparability, and poor repeatability

(3) The lysis of platelets in the method for preparing platelet lysate adopted in the prior art is not complete, so that the relevant factors are not fully released

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