Preparation and application method of human peripheral blood stem cell freezing medium

Abstract

The invention relates to preparation and an application method of a human peripheral blood stem cell freezing medium. The stem cell freezing medium is prepared from the following components in percentage by volume: 48.2% of hydroxyethyl starch dextran, 2% of compound amino acid injection (18AA-IV), 4.8% of dimethyl sulfoxide and 20% of human serum albumin. The stem cell freezing medium has the characteristics of high viable rate after recovery (greater than 99%) and availability for vein back transfusion. The invention further provides the preparation and the application method of the human peripheral blood stem cell freezing medium. The preparation comprises the following steps: under sterile environment, premixing all the components of the human peripheral blood stem cell freezing medium in sequence, adding a stem cell suspension, mixing uniformly, and packing into a cryopreservation bag for cryopreservation. Stem cells with the freezing medium can be directly placed into a cryogenic refrigerator at -80 DEG C for storage, a programmable cooler is not needed, the operation is simple and convenient, and the use is safe.

Description

technical field [0001] The invention belongs to the field of stem cell bioengineering. Specifically, it relates to a method for preparing and using a human peripheral blood stem cell cryopreservation solution. Background technique [0002] Autologous peripheral blood hematopoietic stem cell transplantation has become one of the radical methods for the treatment of malignant hematological diseases, malignant lymphoma, solid tumors and other diseases. The method of autologous peripheral blood stem cell transplantation is to mobilize hematopoietic stem cells into peripheral blood with a hematopoietic stem cell mobilizer, and then use a blood cell separator to separate and collect the mononuclear cell components of the peripheral blood. Since it is an autologous peripheral blood stem cell transplantation, after mobilizing, separating and collecting autologous peripheral blood stem cells, the patient needs to undergo pretreatment such as radiotherapy and chemotherapy for 1 week ...

AI Technical Summary

Problems solved by technology

Although this cryopreservation technique is simple and convenient, and the cryopreservation effect is good, there are still some problems in clinical application: 1. The maintenance time of cell viability is short

Especially for cells frozen with this technology, the cell viability dropped from 92.6% in the first month to 86.2% in the sixth month, with an average monthly drop of about 1%; 2. Because high concentrations of DMSO have certain effects on the human body Toxic and side effects, directly transplanted into patients after resuscitation may cause symptoms such as nausea, vomiting, abdominal cramps, slow heart rate, elevated blood pressure, etc., so 9-10% DMSO concentration is not easy to be directly clinically infused; 3. Because of 1640 culture Because its ingredients are far from the human environment, it has not been approved for clinical use and cannot be directly infused clinically

Therefore, the stem cells cryopreserved by this technology need to be treated with the cryopreservation solution before they can be infused clinically after recovery, which brings inconvenience to clinical application.

However, the classic cryopreservation method uses a programmed cooling device to cool down and freeze, which makes the operation cumbersome, the cost is high, and it is not easy to be used on a large scale in clinical practice.

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