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Cryoprotectant and simple cryopreservation method for hematopoietic stem cells

A cryopreservation protection agent and a technology for hematopoietic stem cells, which are applied in the biological field and can solve the problems of insufficient cryopreservation effect of hematopoietic stem cells.

Active Publication Date: 2019-04-30
北京智能宝生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, 3% DMSO is not as effective for cryopreservation of hematopoietic stem cells as 5%-10% DMSO

Method used

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  • Cryoprotectant and simple cryopreservation method for hematopoietic stem cells
  • Cryoprotectant and simple cryopreservation method for hematopoietic stem cells
  • Cryoprotectant and simple cryopreservation method for hematopoietic stem cells

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Embodiment Construction

[0024] The substantive content of the present invention will be described below in conjunction with specific embodiments, but the protection scope of the present invention cannot be limited thereto.

[0025] 1. Experimental materials

[0026] The mice were 10-week-old healthy male C57BL / 6 mice; PBS buffer was purchased from Shanghai Shushi Biology; Ficoll lymphocyte separation medium was purchased from Tianjin Haoyang Biology; Anti-CD34-PE, Anti-PE MicroBeads, MS sorting The column was purchased from Miltenyi, Germany; the IMDM culture medium and fetal bovine serum were purchased from Hyclone; the BCA kit was purchased from Beyond; the Caspase-3 mouse monoclonal antibody was purchased from Abbkine, and the HRP-labeled secondary antibody was purchased from Solarbio. The purity of Scopadulciol is not less than 95%.

[0027] 2. Experimental method

[0028] 1. Preparation of mouse bone marrow mononuclear cells (BMNCs)

[0029] Mice were sacrificed under aseptic conditions by di...

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Abstract

The invention discloses a cryoprotectant and a simple cryopreservation method for hematopoietic stem cells. The reduction of the content of DMSO to 3% in the prior art significantly reduces the cryopreservation survival rate of the hematopoietic stem cells. It is found that the addition of scoparinol to the cryoprotectant can eliminate cryopreservation damages brought by the reduction of the DMSOcontent, so the scoparinol has a cryoprotective effect on the hematopoietic stem cells, and inhibition of Caspase-3 expression may be a mechanism of the scoparinol exerting the protective effect on the cryopreservation of the hematopoietic stem cells.

Description

technical field [0001] The invention belongs to the biological field and relates to cryopreservation of hematopoietic stem cells, in particular to a cryopreservation protectant and a simple cryopreservation method for hematopoietic stem cells. Background technique [0002] Hematopoietic stem cells (HSCs) exist in very low numbers in mammalian bone marrow, fetal liver, peripheral blood and umbilical cord blood. They are stem cells with high self-renewal ability and multi-directional differentiation potential, and are the precursor cells of all mature blood cells. . Hematopoietic stem cell transplantation is an important method for the treatment of malignant hematological diseases, genetic diseases and immunodeficiency diseases. [0003] The traditional method of using 10% DMSO as a cryoprotectant, programmed cooling, and liquid nitrogen cryopreservation of human hematopoietic stem cells is highly praised for its long-term preservation and less cell loss, and is widely used c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 不公告发明人
Owner 北京智能宝生物科技有限公司
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