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Porcine cells medium

A medium and basic medium technology, applied in the field of cell biology, can solve the problems of lack of medium, the influence of the number of cell passages, and the limited speed of cell proliferation in the cell growth state.

Inactive Publication Date: 2014-04-30
深圳华大基因农业控股有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when pig cells are cultured on this medium, the cell growth state and cell proliferation speed are relatively limited, and the number of passages of cells is also greatly affected.
At present, there is still a lack of ideal and suitable media for pig cell culture

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1: the formula of porcine cell culture medium A, B, C

[0075] 1) The formula of porcine cell culture medium A:

[0076] Pig cell culture medium A consists of artificial medium 1 (the components are shown in Table 4), 5% (v / v) fetal pig serum and factor combination 1 (the components are shown in Table 5).

[0077] Table 4 artificial medium 1 formula:

[0078] name

Content (mg / L)

Calcium Chloride Anhydrous

200.00

L-serine

42.00

L-tryptophan

16.00

L-threonine

95.00

L-tyrosine sodium salt

104.00

Ferric nitrate

0.10

potassium chloride

400.00

L-valine

94.00

L-arginine hydrochloride

84.00

L-Glutamine

584.00

L-cystine hydrochloride

63.00

L-leucine

95.00

L-isoleucine

93.00

L-Histidine Hydrochloride

32.00

L-Lysine Hydrochloride

126.00

L-methionine

25....

Embodiment 2

[0087] Embodiment 2: the formula of porcine cell culture medium D, E, F, G

[0088] 1) The formula of pig cell culture medium D:

[0089] Pig cell culture medium D consists of artificial medium 2 (the components are shown in Table 6), 25% (v / v) fetal pig serum and factor combination 2 (the components are shown in Table 7).

[0090] Table 6 artificial medium 2 formula:

[0091] Compound name

Content (mg / L)

Calcium Chloride Anhydrous

240.00

L-serine

55.00

L-tryptophan

20.00

L-threonine

110.00

[0092] L-tyrosine sodium salt

123.00

Ferric nitrate

0.40

potassium chloride

600.00

L-valine

112.00

L-arginine hydrochloride

98.00

L-Glutamine

628.00

L-cystine hydrochloride

72.00

L-leucine

110.00

L-isoleucine

105.00

L-Histidine Hydrochloride

52.00

L-Lysine Hydrochloride

156....

Embodiment 3

[0101] Example 3: Preparation of primary porcine fibroblasts

[0102] Primary porcine fibroblasts can be prepared by traditional enzymatic digestion, the steps are as follows:

[0103] 1) Quickly scrub the pig fetuses that were killed (within 10 hours after being killed) three times with 70% alcohol, and put them into PBS+double antibody solution;

[0104] 2) Rinse the pig fetus with PBS in the ultra-clean workbench, and remove the head, limbs and internal organs of the pig fetus;

[0105] 3) Mince the pig fetal tissue treated in step 2) in a 60mm diameter plate, transfer the tissue fragments to a 50mL Erlenmeyer flask, and further chop into 1mm 3 Small fragments, take a small amount of tissue fragments and spread them on a 100mm Petri dish, add 10mL of culture medium (DMEM+15%FBS+double antibody), put in CO 2 Cultivate in an incubator (tissue block attachment method); add 3-5mL 0.25% trypsin digestion solution preheated at 39°C to the rest of the tissue fragments, put in CO...

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Abstract

A culture medium of pig cells is provided, which comprises a basic culture medium and fetal pig serum. A method for culturing pig cells, fetal pig serum, and a use of the fetal pig serum in culturing pig cells or preparing the culture medium of pig cells are further provided.

Description

technical field [0001] The invention belongs to the field of cell biology and relates to a pig cell culture medium, which comprises a basic culture medium and fetal boar serum. The invention also relates to a method for cultivating pig cells, a fetal pig serum and the use of the fetal pig serum in cultivating pig cells or preparing pig cell culture medium. Background technique [0002] The medium is not only the basic material for supplying cell nutrition and promoting cell reproduction and proliferation in cell culture, but also the living environment for cell growth and reproduction. There are many types of medium, which can be divided into semi-solid medium and medium according to their material state; they can be divided into synthetic medium and natural medium according to their source. Currently, a complete medium in which natural components are added to a synthetic medium is mainly used. [0003] A complete medium consists of a basal medium (such as MEM) and supplem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/075C12N5/077
CPCC12N5/0018
Inventor 李巍梁颜刘欢
Owner 深圳华大基因农业控股有限公司
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