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Fibroblast culture medium and application thereof

A fibroblast, culture medium technology, applied in animal cells, culture process, tissue culture, etc.

Active Publication Date: 2019-01-01
BIOCELLS BEIJING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although fibroblasts are widely present in the human body, in order to obtain the number of cells required for clinical treatment, large-scale expansion in vitro is still necessary

Method used

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  • Fibroblast culture medium and application thereof
  • Fibroblast culture medium and application thereof
  • Fibroblast culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0032] In a first aspect, the present application provides a cell culture medium comprising a basal medium and a platelet lysate.

[0033] In some embodiments, the cell culture medium further comprises an antibiotic. In some embodiments, the antibiotic is gentamicin, amphotericin B, or a combination thereof.

[0034] In some embodiments, the volume content of the platelet lysate in the cell culture medium is 1-10%.

[0035] In some embodiments, the volume content of the platelet lysate in the cell culture medium is 2-5%, such as 2.5% or 5%.

[0036] In some embodiments, the platelet lysate is a human platelet lysate.

[0037] In some specific embodiments, the platelet lysate is SuperGrow cell culture supplement. SuperGrow cell culture supplements are commercially available.

[0038] In some embodiments, the basal medium is DMEM medium, BME medium, MEM medium, Ham's F12 medium, RPMI1640 medium, 199 medium, IMDM medium or any combination thereof.

[0039] In some embodiment...

Embodiment 1

[0088] The cultivation of embodiment 1 human fibroblast

[0089] 1. Acquisition of Human Fibroblasts

[0090] Put the fresh skin tissue retrieved under aseptic conditions (from hospital medical operation skin tissue waste) into a sterile petri dish, wash with 4°C pre-cooled normal saline for 3 times, and at the same time use sterile ophthalmic surgical scissors Cooperate with tweezers to scrape out the residual blood in the tissue, cut off the excess subcutaneous tissue, and then cut the tissue pieces into pieces (about 1mm in diameter).

[0091] The shredded tissues were mixed and weighed. Weighed 6 parts in total at 0.25 g per part, numbered 1 to 6. After mixing with 5 mL of digestion solution, digested at 37°C for 4 hours to obtain the digested mixture.

[0092] The preparation method of digestive juice is as follows:

[0093] Weigh 0.1 g of collagenase I and add it to 100 mL of high-glucose DMEM medium (Beijing Zhongsheng Aobang Biotechnology Co., Ltd.; Cat. No.: 03.1002...

Embodiment 2

[0098] Characterization of embodiment 2 human fibroblasts

[0099] 1. Detection of cell number and viability

[0100] After the cells in Example 1 were cultured for 3 days, the cell culture medium in the culture dish was replaced with the corresponding fresh cell culture medium, and the growth status of the cells was observed and recorded, and then observed every other day and the growth status of the cells was recorded by taking pictures. Those cultured with cell culture medium A (10% (v / v) FBS+high-glucose DMEM medium) were recorded as the FBS group; those cultured with cell culture medium B (2.5% (v / v) SuperGrow cell culture supplement+high sugar Type DMEM medium) were recorded as SuperGrow group. The result is as figure 1 As shown, the primary cells in the SuperGrow group reached 60% adherence on the 13th day of culture, 2 days earlier than the primary cells in the FBS group (15th day) reached 60% adherence.

[0101] When the cells grow and adhere to a certain density (...

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Abstract

The invention provides a culture medium. The culture medium comprises a basal medium and a platelet lysate, preferably, comprises gutamic acid and / or proline. The invention also provides a method of culturing the fibroblasts using the above cell culture medium, and an application of the cultured fibroblasts as a raw material for cosmetic or tissue repair.

Description

technical field [0001] This application belongs to the field of cell culture. Specifically, the application relates to a fibroblast culture medium and its application. Background technique [0002] Fibroblasts are the main cells of the skin dermis, the largest organ of the human body. They can synthesize and secrete a large amount of collagen and a small amount of elastin, and aggregate outside the cells into thick collagen fiber bundles and fine elastic fiber bundles. Collagen fibers are the main structural protein synthesized and secreted by fibroblasts. They aggregate in the dermis into thick, interwoven fiber bundles that are parallel to the skin surface. They have high mechanical stability and are important for maintaining skin tension and bearing tension. The ingredients are also the material basis for maintaining the plumpness of the skin. [0003] Autologous fibroblast wrinkle removal has become an important technical measure in the field of cosmetology. This tech...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077A61L27/38A61L27/60
CPCA61L27/3804A61L27/3895A61L27/60C12N5/0656C12N2500/32C12N2500/84C12N2509/00
Inventor 刘书磊韩化敏
Owner BIOCELLS BEIJING BIOTECH CO LTD
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