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Culture medium for culturing urine-derived cells

A culture medium and basal medium technology, applied in the direction of cell culture medium, culture process, tissue culture, etc., can solve the problems of death, prone to apoptosis, difficult logarithmic growth phase, etc., and achieve the effect of promoting cell culture

Pending Publication Date: 2018-09-25
OSINGLAY BIO PHARM CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But when the amount of cells is very small, the latent growth period of the cells is prolonged, the proliferation progress is slow, it is difficult to enter the logarithmic growth phase, the cells are extremely difficult to survive, and apoptosis and death are prone to occur.

Method used

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  • Culture medium for culturing urine-derived cells
  • Culture medium for culturing urine-derived cells
  • Culture medium for culturing urine-derived cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] According to Table 1, the different groups of culture media were mixed, and the components other than the basic medium were added first, and then the basic medium was added to make up. Among them, the basic medium included DMEM, DMEM / F12, αMEM, RPMI 1640 , CMRL-1066, Ham's F12, IMDM, 199, MCDB.

[0094] Table 1 Cell culture medium formula without xenogeneic urine source

[0095]

[0096]

[0097]

[0098] Digest primary urine-derived cells and count them. Plate 1500 cells per well (96-well plate). Day0 uses primary urine cell culture medium (REGM medium) for culture, and Day1, Day3, and Day5 are replaced with corresponding groups The medium was cultured, 150 μl of medium per well, and MTT detection was performed on Day1, Day4, and Day7.

[0099] After MTT test, the MTT results of groups 1-25 are as follows: figure 1 shown.

[0100] figure 1 It is the result picture of MTT. The principle of MTT detection is that succinate dehydrogenase in the mitochondria of...

Embodiment 2

[0103] In this example, on the basis of Group 1 in Example 1, the medium was formulated according to Table 2, wherein the volume ratio of platelet lysate to the medium was added according to Table 3 to obtain different groups of medium. When configuring, first add the components other than the basal medium, and then add the basal medium to supplement, wherein the basal medium is DMEM / F12:DMEM=1:1.

[0104] Table 2 Components of culture medium containing platelet lysates in different volume ratios

[0105]

[0106] Table 3 Volume ratio of platelet lysate

[0107]

[0108]

[0109] Digest primary urine-derived cells and count them. Plate 1500 cells per well (96-well plate). Day0 uses primary urine cell culture medium (REGM medium) for culture, and Day1, Day3, and Day5 are replaced with corresponding groups The culture medium was cultured, and the medium volume of each well was 150 μl, and MTT detection was performed on Day1, Day4, and Day7.

[0110] After the MTT test...

Embodiment 3

[0113] According to Table 4, different groups of culture media were mixed, and the components other than the basal medium were added first, and then the basal medium was added for supplementation, wherein the basal medium was DMEM / F12:DMEM=1:1.

[0114] Platelet lysate and various vitamin Cs were added to the medium formula in Table 4, vitamin C including L-ascorbic acid, vitamin C phosphate magnesium.

[0115] Table 4 Formula of cell culture medium without xenogeneic urine source

[0116]

[0117]

[0118] Digest primary urine-derived cells and count them. Plate 1500 cells per well (96-well plate). Day0 uses primary urine cell culture medium (REGM medium) for culture, and Day1, Day3, and Day5 are replaced with corresponding groups The medium was cultured, 150 μl of medium per well; MTT detection was carried out on Day1, Day4, and Day7.

[0119] After the MTT test, the MTT results of groups 1-7 are as follows image 3 shown. From image 3 It can be seen that on Day7,...

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Abstract

The invention discloses a culture medium for culturing urine-derived cells, and belongs to the field of cell culture. The culture medium contains human blood platelet lysates. The culture medium has the advantages that the fact that the human blood platelet lysates can be used as core additives for the heterology-free urine-derived cell culture medium is ultimately confirmed via extensive screening experiments by applicants over the years, and accordingly the problems of extremely low urine-derived cell proliferation speeds, gradual cell death and the like during heterology-free culture in thepast can be solved by the aid of the culture medium; the human blood platelet lysates are added in the heterology-free culture medium, the urine-derived cells can be effectively cultured by the aid of the heterology-free culture medium, and the requirements of research or clinical application on the cell quantities can be met; the blood platelet lysates are innovatively applied to urine-derived cell heterology-free culture and the culture medium, and the culture medium has important significance in regenerative medicine.

Description

technical field [0001] The invention belongs to the field of cell culture, in particular to a culture medium for culturing urine-derived cells. Background technique [0002] In recent years, with the maturity of induced pluripotent stem cell technology, many different types of cells can be successfully induced into pluripotent stem cells, including urine-derived cells. According to literature reports, urine-derived cells were successfully induced into pluripotent stem cells in 2011. Compared with other types of cell extraction methods, urine-derived cells are simpler. You only need to collect the donor’s urine to avoid pain or trauma to the donor. Therefore, urine-derived cells are the best choice for induced pluripotent stem cell donors. ideal source. Since the culture of urine-derived cells is a key step in obtaining induced pluripotent stem cells, the development of a culture technology suitable for clinical-grade urine-derived cells will greatly promote regenerative med...

Claims

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Application Information

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IPC IPC(8): C12N5/00
CPCC12N5/0018C12N2501/81C12N2501/405C12N2501/39C12N2501/415C12N2501/30C12N2501/11C12N2500/25C12N2500/38C12N2500/84
Inventor 王淋立关春燕陈月花李强
Owner OSINGLAY BIO PHARM CO LTD
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