A hemopoietic stem cell cryopreserving solution, and a hemopoietic stem cell cryopreserving method

A technology of hematopoietic stem cells and cryopreservation solution, applied in the field of cell biology, can solve the problem that hematopoietic stem cells cannot be stored and transported for a long time, and achieve the effects of avoiding unstable storage activity, protecting cell membranes, and ensuring physiological functions and biological characteristics.

Inactive Publication Date: 2017-08-29
DONGGUAN BOALAI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a method for cryopreservation of hematopoietic stem cells, which can effectively solve the problem that hematopoietic stem cells cannot be stored and transported for a long time, and realize the ap...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] A hematopoietic stem cell cryopreservation solution, the hematopoietic stem cell cryopreservation solution includes the following raw materials in parts by weight:

[0053] Dimethyl sulfoxide 10mL

[0054] Hydroxyethyl starch 6g

[0055] Cell culture medium 4mL

[0056] Human Serum Albumin 6mL

[0057] Human autologous plasma 60mL.

[0058] The cell culture medium is X-VIVO15 medium.

[0059] The preparation method of the human autologous plasma is as follows: put human peripheral blood in a sterile centrifuge tube, centrifuge at 1500rpm for 8min, take the upper plasma and inactivate it at 56°C for 32min, then transfer it to a new centrifuge tube at 2500rpm Centrifuge at a rotating speed for 15 minutes, and absorb the supernatant, which is the human autologous plasma for cryopreservation.

[0060] The hematopoietic stem cell cryopreservation solution also includes 1 g of sugar, which is a mixture of trehalose, α-1,4-glucan, lentinan and glucose in a weight ratio of...

Embodiment 2

[0068] A hematopoietic stem cell cryopreservation solution, the hematopoietic stem cell cryopreservation solution includes the following raw materials in parts by weight:

[0069] Dimethyl sulfoxide 12mL

[0070] Hydroxyethyl starch 7g

[0071] Cell culture medium 5mL

[0072] Human Serum Albumin 7mL

[0073] Human autologous plasma 70mL.

[0074] The cell culture medium is AIM-V medium.

[0075] The preparation method of the human autologous plasma is as follows: human peripheral blood is placed in a sterile centrifuge tube, centrifuged at 1600rpm for 7min, the upper layer of plasma is inactivated at 57°C for 31min, and then transferred to a new centrifuge tube at 2800rpm Centrifuge at a rotational speed for 22 minutes, and absorb the supernatant, which is the human autologous plasma for cryopreservation.

[0076] The hematopoietic stem cell cryopreservation solution also includes 1.5 g of carbohydrates, which are a mixture of trehalose, α-1,4-glucan, lentinan and glucos...

Embodiment 3

[0084] A hematopoietic stem cell cryopreservation solution, the hematopoietic stem cell cryopreservation solution includes the following raw materials in parts by weight:

[0085] Dimethyl sulfoxide 15mL

[0086] Hydroxyethyl starch 8g

[0087] Cell culture medium 6mL

[0088] Human Serum Albumin 8mL

[0089] Human autologous plasma 80mL.

[0090] The cell culture medium is DMEM / F12 medium.

[0091] The preparation method of the human autologous plasma is as follows: human peripheral blood is placed in a sterile centrifuge tube, centrifuged at 1800rpm for 6min, the upper layer of plasma is inactivated at 58°C for 30min, and then transferred to a new centrifuge tube at 3000rpm Centrifuge at a rotating speed for 20 minutes, and absorb the supernatant, which is the human autologous plasma for cryopreservation.

[0092] The hematopoietic stem cell cryopreservation solution also includes 2 g of carbohydrates, which are a mixture of trehalose, α-1,4-glucan, lentinan and glucose...

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Abstract

The invention relates to the technical field of cytobiology, and particularly relates to a hemopoietic stem cell cryopreserving solution, and a hemopoietic stem cell cryopreserving method. The cryopreserving solution includes following raw materials by weight: 10-20 mL of dimethyl sulfoxide, 6-10 g of hydroxyethyl starch, 4-8 mL of a cell culture medium, 6-10 mL of human serum albumin, and 60-100 mL of human autologous plasma. The formula of the cryopreserving solution is simple, and the raw materials are cheap. The cryopreserving solution can effectively protect hemopoietic stem cells from freezing injuries, is high in safety and low in cell injuries, can increase the resuscitation survival rate of hemopoietic stem cells. The survival rate after resuscitation can be 96% or above, and after resuscitation, the cell viability is high, the quantity of cell proliferation is high, the cells are not liable to age, physiological functions and biological characteristics of the hemopoietic stem cells after resuscitation can be ensured, and microbe detection indexes meet requirements. The survival time of the hemopoietic stem cells is prolonged.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a hematopoietic stem cell cryopreservation solution and a hematopoietic stem cell cryopreservation method. Background technique [0002] Hematopoietic stem cells are adult stem cells in the blood system. They are a heterogeneous group with the ability of long-term self-renewal and the potential to differentiate into various mature blood cells. It is a type of adult stem cell with the longest and most in-depth research history, and it has important guiding significance for the study of various stem cells, including tumor stem cells. Mature cells in the blood system have a very short lifespan. Therefore, in a person's life, hematopoietic stem cells need to replenish various mature cell components of the blood system in a timely manner according to the physiological needs of the body. At the same time, under stress conditions such as injury and inflammation, hematopoietic stem...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 罗天恩
Owner DONGGUAN BOALAI BIOLOGICAL TECH CO LTD
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