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Application of Compounds Containing Single Pentasaccharide Building Blocks in Cell Culture

A structural unit, cell culture technology, applied in the direction of cell culture active agent, animal cells, vertebrate cells, etc., can solve the problems of different molecular weight of heparin, non-uniform activity, difficult quality control, etc., to promote cell proliferation and composition. Clear, nurturing stable effects

Active Publication Date: 2022-07-01
FUTURE HOMO SAPIENS INST OF REGENERATIVE MEDICINE CO LTD (FHSR) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This leads to the obtained heparin with different molecular weights and structures, so there are problems such as uneven activity and difficult quality monitoring.

Method used

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  • Application of Compounds Containing Single Pentasaccharide Building Blocks in Cell Culture
  • Application of Compounds Containing Single Pentasaccharide Building Blocks in Cell Culture
  • Application of Compounds Containing Single Pentasaccharide Building Blocks in Cell Culture

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0101] The preparation method of the IL medium in the following examples: add each component of the medium to a part of the basal medium to dissolve, and continue to add the basal medium to make up to 1L.

[0102] During the experiment, the inventors studied the addition amount of heparin sodium (CAS No.: 9041-08-1), and mainly studied the proliferation performance of cells when the addition amount of heparin sodium in each medium was 100-400ng / mL. The results showed that when the amount of heparin sodium added was 300ng / mL, the proliferation effect was the best. Therefore, in the following examples, the optimal addition amount of heparin sodium is 300 ng / mL as a comparison, and the effect of each compound in the present invention on cell culture is investigated.

Embodiment 1

[0104] Effects of fondaparinux sodium (CAS No.: 104993-28-4) on the proliferation performance of human pluripotent stem cells

[0105] The formulations of human pluripotent stem cell (iPSC) medium in different groups are shown in Table 1. According to the MTT method, the survival rate of human iPSC in each medium was detected, and the results are shown in Table 2 and figure 1 shown.

[0106] Table 1 Composition ratio of pluripotent stem cell culture medium

[0107]

[0108] Table 2 Human iPSC cell viability

[0109]

[0110]

[0111] Remarks: Take the second group as the control, and carry out the calculation of the P value in the independent sample T test.

[0112] from Table 2 and figure 1It can be seen that the proliferation rate of human iPSCs in the medium supplemented with sodium heparin and fondaparinux is much higher than that in the medium without this component. And the proliferation rate of human iPSCs in the medium supplemented with fondaparinux was a...

Embodiment 2

[0114] On the basis of Example 1, the addition amount of fondaparinux sodium was adjusted, and the effect of its concentration on the proliferation rate of human iPSC was investigated.

[0115] The composition of human pluripotent stem cell (iPSC) medium of different groups is shown in Table 3. According to the MTT method, the survival rate of human iPSC in each medium was detected, and the results are shown in Table 4 and figure 2 shown.

[0116] Table 3 Composition ratio of human pluripotent stem cell culture medium

[0117]

[0118]

[0119] Table 4 Human iPSC cell viability

[0120]

[0121] Remarks: Take the second group as the control, and carry out the calculation of the P value in the independent sample T test.

[0122] According to Table 4 and figure 2 It can be seen that in the range of 1-350nM, the cell proliferation is more favorable as the concentration of fondaparinux sodium increases, but when the concentration of fondaparinux sodium is 1nM, the e...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to the application of a compound containing a single pentasaccharide structural unit and a salt thereof in cell culture. The compound has a structure as shown in formula I, wherein the substituents have the meanings given in the specification, which can better control serum-free medium and compound-defined medium, and make the composition more definite, better in quality control, and better in quality. Low infection, more stable culture. The present invention also provides a medium comprising the compound shown in formula I, the medium has the advantages of clear chemical composition, small molecular weight, low cost, convenient quality control, efficient promotion of cell proliferation and maintenance of stem cell pluripotency.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to the application of a compound containing a single pentasaccharide structural unit in cell culture, and a culture medium containing the compound. Background technique [0002] Cell culture technology has developed rapidly in recent years, and it has been widely used in recombinant protein drug preparation, drug screening, and disease research. Animal cells are usually cultured in serum-supplemented medium in order to supply the hormones, growth factors, proteins, and other nutrients necessary for cell growth. However, the composition of serum is complex, with large differences between batches. It contains growth-inhibiting or harmful components, and is easily infected by viruses and mycoplasma. Moreover, the source and production of serum are difficult to monitor. Before use, a large number of experiments must be carried out for verification, which brings many disadvantages to product...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074C12N5/0775C12N5/0797
CPCC12N5/0662C12N5/0623C12N5/0696C12N2501/91
Inventor 王淋立莫健陈月花李强
Owner FUTURE HOMO SAPIENS INST OF REGENERATIVE MEDICINE CO LTD (FHSR)
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