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Porcine mesenchymal stem cell culture solution and preparation method thereof

A technology of mesenchymal stem cells and culture medium, applied in the field of culture medium of porcine umbilical cord and/or placental mesenchymal stem cells, to achieve the effect of high cell purity, strong cell activity, and short passage cycle

Inactive Publication Date: 2018-01-16
TIANJIN AMCELLGENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no proven medium formulation that can effectively culture porcine placenta umbilical cord MSCs

Method used

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  • Porcine mesenchymal stem cell culture solution and preparation method thereof
  • Porcine mesenchymal stem cell culture solution and preparation method thereof
  • Porcine mesenchymal stem cell culture solution and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1. Different factors are compared to the culture situation of cell

[0040] After resuscitating a strain of porcine umbilical cord mesenchymal stem cells, they were cultured in different media, and the concentration of various factors was 5 ng / mL. Because the cultivation of pig cells is difficult and complicated, it does not contain serum but still contains cytokines. The cells hardly grow in the culture medium, so the addition of serum is still necessary.

[0041] Control group:

[0042] (1) Imported fetal bovine serum + basal medium;

[0043](2) Imported fetal bovine serum + basal medium + b-FGF;

[0044] (3) Imported fetal bovine serum + basal medium + h-EGF;

[0045] test group:

[0046] (4) Imported fetal bovine serum + basal medium + PDGF-BB;

[0047] (5) Imported fetal bovine serum + basal medium + b-FGF + PDGF-BB;

[0048] (6) Imported fetal bovine serum + basal medium + bFGF + hEGF;

[0049] (7) Imported fetal bovine serum + basal medium + hEG...

Embodiment 2

[0058] Example 2. Selection of concentration of factors

[0059] Take a porcine umbilical cord mesenchymal stem cell after resuscitating, and culture it with a medium containing three factors of PDGF+bFGF+hEGF at different concentrations. The concentrations of the three factors in each group are the same. The experiment is divided into four groups (1) Control group: imported Fetal bovine serum + basal medium (DMEM / F12 medium); (2) Imported fetal bovine serum + basal medium (DMEM / F12 medium) + 2.5ng / mL three factors, (3) Imported fetal bovine serum + basal Culture medium (DMEM / F12 medium) + 5ng / mL three factors (4) imported fetal bovine serum + basal medium (DMEM / F12 medium) + 10ng / mL three factors; four groups of basal medium (DMEM / F12 Medium) and fetal bovine serum volume ratio is 9:1; in the same planting density (2.5*10 5 each / 5mL) under the same culture conditions (37°C, CO 2 The volume concentration is 5%) under planting T25cm 2 After culturing for three days in the cu...

Embodiment 3

[0065] Example 3. Detection of passage ability of porcine umbilical cord placental mesenchymal stem cells

[0066] Two different lines of porcine umbilical cord mesenchymal stem cells P2 were recovered at the same density, using the following two culture media, (1) control group: imported fetal bovine serum + basal medium; (2) factor group: imported fetal bovine serum Bovine serum + basal medium + 5ng / mL PDGF-BB + bFGF + hEGF, the volume ratio of the basal medium (DMEM / F12 medium) and fetal calf serum in the control group and the experimental group is 9:1, and the other factors are the same Cultured under certain culture conditions, the cells were passaged separately, and the passage ability of the cells was tested. The experimental results are shown in image 3 .

[0067] Apoptosis is the natural death and aging phenomenon of cells in the growth process. After different strains of porcine umbilical cord mesenchymal stem cells were cultured with factor-added culture medium, ...

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Abstract

The invention relates to a porcine mesenchymal stem cell culture solution and a preparation method thereof. The culture solution is prepared from the following components: a basal culture medium freeof animal-derived components, 2-20% of fetal calf serum and 5-10ng / mL of PDGF-BB factors. Compared with the conventional common mesenchymal stem cell culture solution, the porcine umbilical cord placenta mesenchymal stem cells cultured by the porcine mesenchymal stem cell culture solution have more typical cellular morphology, have higher cell passage capability, and are low in possibility of senescence; when the cells cultured by the porcine umbilical cord placenta mesenchymal stem cell culture solution is applied to production, porcine umbilical cord placenta mesenchymal stem cell products with higher quantity and more stable biological characteristics can be obtained.

Description

technical field [0001] The invention relates to the technical field of porcine umbilical cord and / or placental cell culture, in particular to a culture solution of porcine umbilical cord and / or placental mesenchymal stem cells. Background technique [0002] Before stem cell therapy was applied to humans, the therapeutic effects of mesenchymal stem cells (MSCs) from various animals, such as mice, cows, rabbits and pigs, on different diseases have been extensively studied. Among them, pigs have many similarities with humans, such as the size, physiology and function of organs. Therefore, pigs have been considered as valuable model animals for in vivo studies and preclinical evaluations. [0003] Porcine MSCs have been successfully isolated from multiple organ tissues, such as bone marrow, skin, cord blood, umbilical cord, endometrium, amniotic fluid and peripheral blood. Porcine bone marrow MSCs show great potential in in vitro differentiation and transdifferentiation to mes...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 贾红红韩之波赵钦军
Owner TIANJIN AMCELLGENE ENG
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