Culture medium for Langerhans precursor cells

Abstract

The invention provides a culture medium for Langerhans precursor cells. The culture medium for Langerhans precursor cells comprises a basal culture medium, GM-CSF, TGF-beta, IL-1 beta, IL-3, IL-6, SCF, EPO and IL-4. According to the culture medium for Langerhans precursor cells, effective culture conditions are provided for obtaining a large number of Langerhans cells, the yield of the Langerhanscells is increased, and the problem that the quantity of seed cells is limited due to the fact that the Langerhans cells cannot be proliferated and subcultured in vitro is effectively solved.

Description

technical field

[0001] The invention belongs to the technical field of tissue engineering biological materials, in particular to a culture medium for Langerhans precursor cells. Background technique

[0002] The skin is not only the body's first line of defense against external biological, physical and chemical damage, but also an important immune organ of the body, participating in the immune response and giving the body a stable internal environment. The epidermis contains about 80% of keratinocytes, melanocytes and Langerhans cells (LC). Langerhans cells are a subset of dendritic cells that reside in the skin, are important antigen-presenting cells in the skin immune response, and play a very key role in regulating the skin immune response. There are specific Berbeck particles, which have the ability to recognize, uptake, process and present exogenous antigens, and migrate to regional lymph nodes, stimulate naive T cell activation, initiate skin immune responses, and fig...

Examples

Embodiment 1

[0113] This embodiment provides a medium for Langerhans precursor cells, the medium for Langerhans precursor cells comprises: RPMI1640 containing 10% fetal bovine serum, 50ng / mL GM-CSF, 5ng / mL TGF-β, 1 ng / mL IL-1β, 10 ng / mL IL-3, 10 ng / mL IL-6, 1 ng / mL SCF, 1 U / mL EPO, and 10 ng / mL IL-4.

[0114] The preparation method of the culture medium of the Langerhans precursor cells of the present embodiment is as follows:

[0115] In a sterile ultra-clean workbench, in a volume ratio of 1:9, fetal bovine serum and RPM1640 medium were mixed well, and then 50ng / mL GM-CSF, 5ng / mL TGF-β, 1ng / mL IL- 1β, 10ng / mL IL-3, 10ng / L IL-6, 1ng / mL SCF, 1U / mL EPO, and 10ng / mL IL-4 were mixed by pipetting, and filtered with a 0.22 μm pore size microporous membrane under positive pressure. bacteria.

Embodiment 2

[0117]This embodiment provides a culture medium for Langerhans precursor cells, which comprises: RPMI1640 containing 10% fetal bovine serum, 500ng / mL GM-CSF, 50ng / mL TGF-β, 100 ng / mL IL-1 β, 100 ng / mL IL-3, 100 ng / mL IL-6, 50 ng / mL SCF, 10 U / mL EPO, and 100 ng / mL IL-4.

[0118] The preparation method of the culture medium of Langerhans precursor cells in this embodiment is as follows:

[0119] In a sterile ultra-clean workbench, mix fetal bovine serum and RPM1640 medium evenly at a volume ratio of 1:9, and then add 500ng / mL GM-CSF, 50ng / mL TGF-β, 100ng / mL IL- 1β, 100ng / mL IL-3, 100ng / LIL-6, 50ng / mL SCF, 10U / mL EPO and 100ng / mL IL-4, pipette and mix well, use a microporous filter membrane with a pore size of 0.22μm to sterilize by positive pressure filtration .

Embodiment 3

[0121] This embodiment provides a culture medium for Langerhans precursor cells, which comprises: RPMI1640 containing 10% fetal bovine serum, 200ng / mL GM-CSF, 25ng / mL TGF-β, 50ng / mL IL-1β, 55ng / mL IL-3, 50ng / mL IL-6, 25ng / mL SCF, 6U / mL EPO, and 60ng / mL IL-4.

[0122] The preparation method of the culture medium of Langerhans precursor cells in this embodiment is as follows:

[0123] In a sterile ultra-clean workbench, mix fetal bovine serum and RPM1640 medium evenly at a volume ratio of 1:9, and then add 200ng / mL GM-CSF, 25ng / mL TGF-β, 50ng / mL IL- 1β, 55ng / mL IL-3, 50ng / L IL-6, 25ng / mL SCF, 6U / mL EPO and 60ng / mL IL-4, mixed by blowing and blowing, and filtered through a microporous filter membrane with a pore size of 0.22μm bacteria.

technical field

[0001] The invention belongs to the technical field of tissue engineering biological materials, in particular to a culture medium for Langerhans precursor cells. Background technique

[0002] The skin is not only the body's first line of defense against external biological, physical and chemical damage, but also an important immune organ of the body, participating in the immune response and giving the body a stable internal environment. The epidermis contains about 80% of keratinocytes, melanocytes and Langerhans cells (LC). Langerhans cells are a subset of dendritic cells that reside in the skin, are important antigen-presenting cells in the skin immune response, and play a very key role in regulating the skin immune response. There are specific Berbeck particles, which have the ability to recognize, uptake, process and present exogenous antigens, and migrate to regional lymph nodes, stimulate naive T cell activation, initiate skin immune responses, and fig...

Examples

Embodiment 1

[0113] This embodiment provides a medium for Langerhans precursor cells, the medium for Langerhans precursor cells comprises: RPMI1640 containing 10% fetal bovine serum, 50ng / mL GM-CSF, 5ng / mL TGF-β, 1 ng / mL IL-1β, 10 ng / mL IL-3, 10 ng / mL IL-6, 1 ng / mL SCF, 1 U / mL EPO, and 10 ng / mL IL-4.

[0114] The preparation method of the culture medium of the Langerhans precursor cells of the present embodiment is as follows:

[0115] In a sterile ultra-clean workbench, in a volume ratio of 1:9, fetal bovine serum and RPM1640 medium were mixed well, and then 50ng / mL GM-CSF, 5ng / mL TGF-β, 1ng / mL IL- 1β, 10ng / mL IL-3, 10ng / L IL-6, 1ng / mL SCF, 1U / mL EPO, and 10ng / mL IL-4 were mixed by pipetting, and filtered with a 0.22 μm pore size microporous membrane under positive pressure. bacteria.

Embodiment 2

[0117]This embodiment provides a culture medium for Langerhans precursor cells, which comprises: RPMI1640 containing 10% fetal bovine serum, 500ng / mL GM-CSF, 50ng / mL TGF-β, 100 ng / mL IL-1 β, 100 ng / mL IL-3, 100 ng / mL IL-6, 50 ng / mL SCF, 10 U / mL EPO, and 100 ng / mL IL-4.

[0118] The preparation method of the culture medium of Langerhans precursor cells in this embodiment is as follows:

[0119] In a sterile ultra-clean workbench, mix fetal bovine serum and RPM1640 medium evenly at a volume ratio of 1:9, and then add 500ng / mL GM-CSF, 50ng / mL TGF-β, 100ng / mL IL- 1β, 100ng / mL IL-3, 100ng / LIL-6, 50ng / mL SCF, 10U / mL EPO and 100ng / mL IL-4, pipette and mix well, use a microporous filter membrane with a pore size of 0.22μm to sterilize by positive pressure filtration .

Embodiment 3

[0121] This embodiment provides a culture medium for Langerhans precursor cells, which comprises: RPMI1640 containing 10% fetal bovine serum, 200ng / mL GM-CSF, 25ng / mL TGF-β, 50ng / mL IL-1β, 55ng / mL IL-3, 50ng / mL IL-6, 25ng / mL SCF, 6U / mL EPO, and 60ng / mL IL-4.

[0122] The preparation method of the culture medium of Langerhans precursor cells in this embodiment is as follows:

[0123] In a sterile ultra-clean workbench, mix fetal bovine serum and RPM1640 medium evenly at a volume ratio of 1:9, and then add 200ng / mL GM-CSF, 25ng / mL TGF-β, 50ng / mL IL- 1β, 55ng / mL IL-3, 50ng / L IL-6, 25ng / mL SCF, 6U / mL EPO and 60ng / mL IL-4, mixed by blowing and blowing, and filtered through a microporous filter membrane with a pore size of 0.22μm bacteria.

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