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Construction method of immune cell bank

A technology of immune cell bank and construction method, which is applied in the field of immune cell therapy, which can solve the problems of decreased cell survival rate, decreased activity, inability to realize long-term storage and instant application, and achieve the effect of improving resource utilization and avoiding waste of resources

Inactive Publication Date: 2016-04-27
上海润泉生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is to separate the immune cells and freeze them separately, code them into the bank, however, after these immune cells are frozen, their activity is greatly reduced, and when these immune cells are resuscitated, the survival rate of the cells is greatly improved. Decrease, unable to achieve long-term storage and immediate application, let alone used as seed cell cycle expansion

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0070]In Example 1, the mononuclear cells were expanded and cultured in vitro for 14 days. The multiplication factor of NK cells can reach more than 10,000 times, and the activity of the prepared NK cells can reach more than 85%. Flow cytometry detects CD16(+) and CD56(+) is greater than 80%; in addition, the NK cells prepared in Example 1 have strong activity and can be stored for a long time, and the 5-year storage cell survival rate is greater than 95%.

Embodiment 2

[0071] Example 2 Placental Tissue NK Immune Cell Bank Construction Method

[0072] (1) For the physical examination of the placental mother, there should be no infectious diseases. After the placenta is collected and transported to the laboratory, further pathogen inspection will be carried out. If it meets the quality requirements for umbilical cord blood storage, it can be used as raw material for storage and preparation of donors.

[0073] (2) Isolation of mononuclear cells from placental tissue: mash the placental tissue, mince the tissue to a diameter of less than 0.1 cm, digest it with 0.25M trypsin at 37°C for 30 min, filter it, and add 1:1 lymphocyte separation solution ( Density 1.077), centrifuged at 600G for 30 minutes, after centrifugation, the middle layer cells were taken to obtain placental tissue mononuclear cells.

[0074] (3) Cryopreservation of mononuclear cells: the obtained umbilical cord blood mononuclear cells were divided into 1.5×10 8 / mL density, mix...

Embodiment 4

[0084] Example 4 Placental tissue T immune cell library construction method

[0085] (1) For the physical examination of the placental mother, there should be no infectious diseases. After the placenta is collected and transported to the laboratory, further pathogen inspection will be carried out. If it meets the quality requirements for umbilical cord blood storage, it can be used as raw material for storage and preparation of donors.

[0086] (2) Isolation of mononuclear cells from placental tissue: mash the placental tissue, mince the tissue to a diameter of less than 0.1 cm, digest it with 0.25M trypsin at 37°C for 30 min, filter it, and add 1:1 lymphocyte separation solution ( Density 1.077), centrifuged at 600G for 30 minutes, after centrifugation, the middle layer cells were taken to obtain placental tissue mononuclear cells.

[0087] (3) Cryopreservation of mononuclear cells: the obtained umbilical cord blood mononuclear cells were divided into 1.5×10 8 / mL density, m...

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Abstract

The invention relates to a construction method of an immune cell bank. The construction method comprises the following steps: processing placenta blood or placenta tissue obtained from umbilical cord blood, peripheral blood and placenta tissue so as to obtain mononuclear cells; conducting amplification culture on the mononuclear cells, or conducting amplification culture on the mononuclear cells after freezing the mononuclear cells; and coding the obtained immune cells and storing the immune cells. The construction method disclosed by the invention, on the basis of achieving the efficient amplification of the immune cells, can still keep the frozen immune cells at high activity and high purity; moreover, the resuscitation activity of the stored immune cells achieves above 85%, the immune cells are resurrected within 30-60min and the storage duration is 15-20 years; the application scope of blood storage and tissue storage to the treatment by virtue of three categories of medical technologies is enhanced, and a novel and healthy storage mode is provided for the prevention and the treatment of tumors and for the prevention and the treatment of immune system diseases caused by virus infection.

Description

technical field [0001] The invention relates to the field of immune cell therapy, in particular to a method for constructing an immune cell bank. Background technique [0002] Immune cells refer to all cells related to immune cell response, mainly including T cells, B cells, natural killer cells (NK), monocytes, etc. In recent years, the immunotherapy technology of tumors and viruses has entered into clinical application, and the adoptive immunotherapy of T cells and NK cells has been included in the catalog of three types of medical technologies in my country, mainly through in vitro stimulation and expansion of autologous immune cells. Improve immunity in patients. By directly or indirectly killing tumor cells or virus-infected cells, the purpose of alleviating or treating diseases is achieved. At the same time, through scale expansion and storage, it can realize rapid integration with applications and improve application effects. [0003] NK cells (natural killer cells,...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12N5/0783A01N1/02
CPCC12N5/0646A01N1/021C12N5/0636
Inventor 刘军唐欢
Owner 上海润泉生物技术有限公司
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