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Detection method of promoted cell proliferation rate of freeze-dried powder

A detection method and cell proliferation technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc., and can solve problems such as increased osmotic pressure and inability to accurately detect the cell proliferation rate of freeze-dried powder , to achieve the effect of overcoming accurate detection

Inactive Publication Date: 2016-05-04
SHAANXI AMY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since the lyophilized powder needs to be added with a lyoprotectant during the freeze-drying process, after the lyophilized powder is added to the culture medium to obtain the sample culture medium, compared with the control medium, the osmotic pressure is greatly increased, and the freeze-dried powder cannot be lyophilized. Accurately detect the cell proliferation promoting rate of the powder

Method used

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  • Detection method of promoted cell proliferation rate of freeze-dried powder
  • Detection method of promoted cell proliferation rate of freeze-dried powder
  • Detection method of promoted cell proliferation rate of freeze-dried powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] 1. Preparation of sample medium and control medium:

[0078] Preparation of sample medium:

[0079] The concentrated freeze-dried powder was dissolved in DMEM medium, and diluted to a protein concentration of 100ppm, filtered and sterilized with a 0.2 μm microporous membrane, and 10% serum was added to obtain a sample medium, and the obtained The sample culture medium was stored at 4°C.

[0080] Preparation of control medium:

[0081] Add 0.5%, 0.92%, 1.5%, 2%, 3%, 5%, 6%, 8% and 10% mannitol to the DMEM medium respectively, and measure the corresponding osmotic pressure values, see Table 1 for details , with the amount of mannitol added as the abscissa and the osmotic pressure as the ordinate to draw a standard curve, see figure 2 , obtain the functional relationship between the osmotic pressure and the addition amount X of mannitol: Y=5425.8X+336.87; Dissolving a certain amount of mannitol in the DMEM medium makes the osmotic pressure of the DMEM medium the same a...

Embodiment 2

[0097] 1. Preparation of sample medium and control medium:

[0098] Preparation of sample medium:

[0099] Dissolve the lyophilized powder of the permeate in DMEM medium, and dilute to a protein concentration of 50ppm, filter and sterilize with a 0.2 μm microporous membrane, add 20% serum to it to obtain a sample medium, and measure the obtained The osmotic pressure of the sample medium was 615 mOsm / kg, and the sample medium was stored at 4°C.

[0100] Preparation of control medium:

[0101] according to figure 2 The functional relationship between the medium osmotic pressure Y and the amount of mannitol added X: Y=5425.8X+336.87; dissolving 5.13% mannitol in the DMEM medium makes the osmotic pressure of the DMEM medium identical to the sample medium, Filter sterilization was performed with a 0.2 μm microporous membrane, 20% serum was added thereto to obtain a control medium, and the control medium was stored at 4°C.

[0102] 2. Cultivate fibroblasts with the obtained sam...

Embodiment 3

[0114] 1. Preparation of sample medium and control medium:

[0115] Preparation of sample medium:

[0116] Dissolve the lyophilized powder of the permeate in DMEM medium, and dilute to a protein concentration of 100ppm, filter and sterilize with a 0.2 μm microporous membrane, add 15% serum to it to obtain a sample medium, and measure the obtained The osmotic pressure of the sample culture medium was 887 mOsm / kg, and the sample culture medium was stored at 4°C.

[0117] Preparation of control medium:

[0118] according to figure 2 The functional relationship between the medium osmotic pressure Y and the amount of mannitol added X: Y=5425.8X+336.87; dissolving 10.14% mannitol in the DMEM medium makes the osmotic pressure of the DMEM medium identical to the sample medium, A 0.2 μm microporous filter membrane was used for filter sterilization, 15% serum was added thereto to obtain a control medium, and the control medium was stored at 4°C.

[0119] 2. Cultivate fibroblasts wi...

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Abstract

The invention relates to the technical field of tissue engineering, and in particular relates to a detection method of a promoted cell proliferation rate of freeze-dried powder. The method can be used for accurately detecting the promoted cell proliferation rate of freeze-dried powder. The embodiments of the invention provide a detection method of the promoted cell proliferation rate of freeze-dried powder. The detection method comprises the following steps: 1) dissolving to-be-detected freeze-dried powder in a complete culture medium to obtain a sample culture medium which has a first osmotic pressure; 2) by taking the complete culture medium without the to-be-detected freeze-dried powder as a control medium, adjusting the osmotic pressure of the control medium to be equal to the first osmotic pressure; 3) carrying out culture on the same number of cells under the same condition by using the obtained sample culture medium and the control medium respectively, and obtaining sample cells and control cells after a first preset time; and 4) carrying out MTT (methylthiazolyldiphenyl-tetrazolium bromide) dyeing and light absorption value detection on the obtained sample cells and control cells, and comparing the light absorption values of the sample cells and the control cells to obtain the promoted cell proliferation rate of a sample in respect to a control.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a method for detecting cell proliferation rate promoted by freeze-dried powder. Background technique [0002] The freeze-dried powder contains biologically active factors. When the activity of the biologically active factors in the freeze-dried powder needs to be detected, the freeze-dried powder can be added to the culture medium to prepare a sample medium, and cultured with the control without adding the freeze-dried powder Base as a reference, the fibroblasts are cultured for the same time, so as to detect the proliferation of fibroblasts under the action of the reference substance and the sample, and the cell proliferation rate of the sample relative to the reference substance can be obtained, so that the freeze-dried Indirect characterization of the biological activity of the biologically active factors in the powder. [0003] Since the lyophilized powder needs t...

Claims

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Application Information

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IPC IPC(8): C12Q1/02G01N21/31
CPCC12Q1/02G01N21/31
Inventor 刘莹南艳萍朱进喜张爱兵
Owner SHAANXI AMY BIOTECH CO LTD
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