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Culture medium for culturing olfactory ensheathing cells

A technology of olfactory ensheathing cells and culture medium, which can be used in animal cells, nervous system cells, vertebrate cells, etc., and can solve problems such as immune response, patient exposure, and impact.

Inactive Publication Date: 2013-03-06
北京清美联创干细胞科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the use of bovine serum to culture human OECs for cell therapy will not only expose patients to the risk of zoonotic diseases, but may also release bovine protein due to the death of transplanted OECS, causing the body's immune response, resulting in serum sickness or other immune disorders. disease
In addition, the activity of each batch of serum is different, which has a certain impact on the stability of cells harvested between batches

Method used

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  • Culture medium for culturing olfactory ensheathing cells
  • Culture medium for culturing olfactory ensheathing cells
  • Culture medium for culturing olfactory ensheathing cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Embodiment 1, the cultivation of olfactory ensheathing cells in different mediums

[0112] 1. Culture of olfactory ensheathing cells

[0113] 1) Pretreatment

[0114] The isolated human olfactory ensheathing cells (Olfactory Ensheathing Cells, OECs, described in Lim F, Martín-Bermejo MJ, García-Escudero V, Gallego-Hernández MT, García-Gómez A, Rábano A, Díaz-Nido J, Avi la J, Moreno-Flores MT. Reversibly immortalized human olfactory ensheathing glia from an elderly donor maintain neuroregenerative capacity. Glia. 2010, 58(5): 546-58. Publicly available from Beijing Qingmei Lianchuang Stem Cell Technology Co., Ltd. .) Wash, digest with trypsin solution at 37.5°C for 3 minutes, collect the cells, centrifuge, discard the supernatant, and collect the cells.

[0115] PBS washing solution, Invitrogen, catalog number 10010031;

[0116] Tryptic Digest, Invitrogen, Cat. No. 25300054.

[0117] 2) Primary culture

[0118] The cells obtained in step 1) are fully blown into a s...

Embodiment 2

[0171] Embodiment 2, the cultivation of olfactory ensheathing cells in different mediums

[0172] 1. Culture of olfactory ensheathing cells

[0173] 1) pretreatment: same as embodiment 1;

[0174] 2) Primary culture: basically the same as in Example 1, the difference is that the cell culture medium 2 is used for culturing;

[0175] 3) Subculture: basically the same as in Example 1, the difference is that the cell culture solution 2 is used for culturing;

[0176] Cell culture solution 1 was prepared according to the following steps: DMEM medium (Invitrogen, 21041025), biotin, glutamine, basic fibroblast growth factor, epidermal growth factor, insulin, selenium, human albumin, tissue Component 1, component 2, component 3, component 4, component 5, component 6 and component 7 are mixed to obtain cell culture medium 1;

[0177] The component 1 is ferric chloride, chromium chloride, cupric chloride, manganese chloride, zinc chloride and sodium molybdate;

[0178] The component...

Embodiment 3

[0224] Embodiment 3, the cultivation of olfactory ensheathing cells in different mediums

[0225] 1. Culture of olfactory ensheathing cells

[0226] 1) pretreatment: same as embodiment 1;

[0227] 2) Primary culture: basically the same as in Example 1, the difference is that the cell culture solution 3 is used for culturing;

[0228] 3) Subculture: basically the same as in Example 1, the difference is that the cell culture solution 3 is used for culturing;

[0229] Cell culture medium 3 was prepared according to the following steps: DMEM medium (Invitrogen, 21041025), biotin, glutamine, basic fibroblast growth factor, epidermal growth factor, insulin, selenium, human albumin, tissue Component 1, component 2, component 3, component 4, component 5, component 6 and component 7 are mixed to obtain cell culture medium 1;

[0230] The component 1 is ferric chloride, chromium chloride, cupric chloride, manganese chloride, zinc chloride and sodium molybdate;

[0231] The component...

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Abstract

The invention discloses a culture medium for culturing olfactory ensheathing cells. The culture medium provided by the invention comprises components 1-10. Proved by experiments, the culture medium provided by the invention is used for culturing olfactory ensheathing cells of primates, and basically has no feeder cells; the olfactory ensheathing cells obtained through culturing are high in proliferation index; and meanwhile, the proficiency of stem cells differentiating into entoderm, mesoderm and ectoderm tissue derivatives is maintained, and the karyotype of the stem cells is maintained.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a medium for cultivating olfactory ensheathing cells. Background technique [0002] Olfactory ensheathing cells are one of the very few cells found in the central nervous system that can regenerate. It is characterized by the ability to regenerate nerves throughout life, and can also release a variety of neurotrophic factors and nerve adhesion molecules. The strongest glial cells, which are increasingly used to treat spinal cord injuries. OECs share phenotypes with glial cells and Schwann cells. They can both promote axon regeneration. The main difference is that OECs exist not only in the central nervous system, but also in the peripheral nerves. Neurons in the olfactory mucosa are the only neurons that grow after birth and continue to differentiate in adults. The life span is 4 to 12 weeks. With the growth of new cells, new innervation relationships are established. Olfactory ens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
Inventor 田杰
Owner 北京清美联创干细胞科技有限公司
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