Cell culture medium for substituting serum by sericin

A cell culture and culture medium technology, applied in animal cells and other directions, can solve the problems of high serum price, easy to carry viruses, poor batch stability, etc., and achieve the effects of reducing costs, avoiding pollution, and improving stability.

Active Publication Date: 2015-02-18
嘉兴欣贝莱生物科技有限公司
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved in the present invention is to provide an animal cell culture additive that can replace serum and a cell culture medium containing the additive in view of the shortcomings of animal cell culture serum that is expensive, easy to carry viruses, and poor in batch stability.

Method used

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  • Cell culture medium for substituting serum by sericin
  • Cell culture medium for substituting serum by sericin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 prepares sericin solution

[0027] Sericin with a purity of more than 92% is dissolved in water or PBS buffer to obtain a sericin stock solution with a concentration greater than 100 mg / ml. The stock solution is used after being sterilized by conventional high temperature sterilization method or sterilized by 0.2 μm filter.

Embodiment 2

[0028] Example 2 MTT method for measuring the effect of sericin on cell proliferation

[0029] Experimental conditions: cell AGS (human gastric cancer cell line, purchased from Sigma, product number 89090402) ( figure 1 ) and cell LCL (human lymphoblastoid cells, purchased from the Cell Bank of Kunming Institute of Zoology, Chinese Academy of Sciences) ( figure 2 ), cultured in different media, the cell culture conditions are: 37°C, 5% carbon dioxide. After culturing for 24 hours and 48 hours, the proliferation was measured, and the results are shown in figure 1 and figure 2 . The media used are shown in a-h below. Among them, FBS is fetal bovine serum, GIBCO is a commercial brand, and RPMI 1640 is a commercial basal salt medium (hereinafter referred to as 1640). A total of three batches of sericin were tested. The first batch was extracted from silkworm, the second batch was extracted from tussah silkworm, and the third batch was extracted from castor silkworm. The ext...

Embodiment 3

[0039] The experimental conditions were the same as in Example 2, and the above-mentioned sericin stock solution was directly added to the cell culture medium (10% FBS+MEM and I-MDM), so that the final concentration of sericin was within a certain range. Human cervical cancer cell line (Hela), human T lymphocyte cell line (H9), human amniotic membrane cell line (Wish), Chinese hamster ovary cell line (CHO-K1), human breast cancer cell line (MDA-MB-231), Human lung cancer cell line (SPC-A1) and porcine kidney cell line (PK-15). Placed in a normal environment for cultivation (37°C, 5% carbon dioxide). As a result, sericin at a final concentration of 10-500 μg / ml was able to promote cell growth.

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Abstract

The invention discloses new use of cell culture medium additive for substituting serum by sericin and a cell culture medium containing sericin to substitute serum; the cell culture medium disclosed by The invention comprises a basic culture medium and nutritional additive; the nutritional additive is 20-2000mug / ml of sericin; in traditional cell cultivation, bovine serum is added normally; but most of the bovine serum is imported and has high price; mass stability between batches is poor; high-temperature sterilization cannot be processed; serious virus such as mycoplasma, bovine spongiform encephalopathy and the like may be contained; and the bovine serum has significant risks to the cell cultivation. According to The invention, the serum is substituted by the sericin; cost is reduced greatly, stability of a cell cultivation system is enhanced, and pollution caused by serum is avoided; in addition, the cell cultivation medium additive disclosed by The invention has significant application value.

Description

technical field [0001] The invention belongs to the field of cell culture medium, in particular to a cell culture medium in which sericin replaces serum. Background technique [0002] Bovine serum is the most used natural medium in cell culture. It is rich in nutrients necessary for cell growth and has extremely important functions. Serum provides hormones that drive exponential cell growth, nutrients that are absent or present in minimal amounts in the basal medium, and low molecular weight nutrients. Fetal bovine serum is obtained by heart puncture, and newborn calf (10-14 days) and calf (within 10 months) serum is obtained from veins. Serum collection conditions must meet the standards of industrial production: collection at low temperature, one-time separation, and mixing and freezing immediately after separation. When preparing bovine serum, it first needs to be heat-inactivated, and the temperature is strictly controlled in a 56°C water bath for 30 minutes to remove ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/07
Inventor 董扬王文段子渊王金焕陈垒
Owner 嘉兴欣贝莱生物科技有限公司
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