CN115466724A - Screening method and application of optimal factor value in human mesenchymal stem cell amplification - Google Patents
Screening method and application of optimal factor value in human mesenchymal stem cell amplification Download PDFInfo
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- 230000003321 amplification Effects 0.000 title claims abstract description 29
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- 230000010261 cell growth Effects 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
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- 238000002637 fluid replacement therapy Methods 0.000 claims description 2
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- 238000003756 stirring Methods 0.000 description 13
- 210000001185 bone marrow Anatomy 0.000 description 3
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Abstract
The invention discloses a screening method of an optimal factor value for human mesenchymal stem cell amplification, which comprises the steps of screening factors, and selecting the factors which can have obvious effect on the amplification of the human mesenchymal stem cells; carrying out single-factor variance analysis on the screened factors, and judging which factors are significance factors according to variance analysis results; carrying out variance analysis on the single factor in the step S2, and judging whether the single factor cannot be fitted into a straight line according to a regression equation; if not, returning to the step S1; if the fitting can be a straight line, performing a climbing experiment to find an interval where the optimal value is located; selecting 4 significant factors and intervals in step S4, and performing RSM analysis; performing experimental design by using a BBD method, and obtaining data of amplification multiples and cell viability of all groups; carrying out ANOVA analysis on the data obtained in the step S6 to obtain a regression equation and a design space; and respectively calculating the optimal factor values of the amplification times and the cell viability rate.
Description
Technical Field
The invention relates to the technical field of stem cell culture, in particular to a screening method and application of an optimal factor value in human mesenchymal stem cell amplification.
Background
Cell therapy has become one of the most active in the field of biological therapy, and the study of standardized cell therapy has become an important component of modern medicine. The cell types commonly used in the current cell therapy in China are cells of adult origin, such as immune cells, bone marrow stem cells, umbilical cord blood and hematopoietic or mesenchymal stem cells of umbilical cord origin.
Mesenchymal Stem Cells (MSC) have wide sources, are easy to separate, culture, amplify and purify, have low immunogenicity, and have no limitation of ethical problems, so the MSC has huge application prospects in the aspects of immunotherapy and gene therapy. Such as autologous or allogeneic bone marrow or cord blood hematopoietic stem cell transplantation for treating hematopathy, malignant tumor, etc.; or the mesenchymal stem cells from the bone marrow and the umbilical cord are transplanted and then transformed into liver-like cells, osteoblasts, cardiac muscle cells, sweat gland cells and the like.
In the report of the existing preparation method of the mesenchymal stem cell factor, for example, the human mesenchymal stem cell factor and the preparation method thereof disclosed in the chinese patent application CN105543313A, the supernatant when the P1-P5 generation cells grow to the confluence of 75-85% is collected and combined, and the supernatant is filtered and ultrafiltered to obtain the cytokine concentrated solution.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a screening method and application of an optimal factor value for human mesenchymal stem cell amplification.
In order to achieve the purpose, the invention adopts the following technical scheme:
a screening method for optimal factor values in human mesenchymal stem cell expansion, the method comprising the steps of:
screening the S1 factor, and selecting the factor which possibly has a remarkable effect on the amplification of the human mesenchymal stem cells;
s2, carrying out single-factor variance analysis on the screened factors, and judging which factors are significant factors according to variance analysis results;
s3, carrying out variance analysis on the single factor in the step S2, and judging whether the single factor cannot be fitted into a straight line according to a regression equation; if not, returning to the step S1;
s4, if the fitting can be a straight line, performing a climbing experiment to find an interval where the optimal value is located;
s5, selecting 4 significant factors and intervals in the step S4, and carrying out RSM analysis;
s6, carrying out experimental design by using a BBD method, and obtaining data of amplification multiples and cell viability of all groups;
s7, performing ANOVA analysis on the data obtained in the step S6 to obtain a regression equation and a design space;
s8, respectively calculating the optimal factor values of the amplification multiple and the cell viability rate.
The factors screened in the step S2 are 8, and are respectively temperature, pH, dissolved oxygen, rotating speed, cell inoculation density, microcarrier inoculation density, culture time and culture medium fluid replacement strategy.
It should be noted that the 4 significant factors and intervals in step S5 are rotation speed, cell inoculation density, microcarrier inoculation density and culture time, respectively.
The rotating speed is 20-40rpm, the cell inoculation density is 40000-120000cells/mL, the microcarrier inoculation density is 0.8-2.4g/L, and the culture time is 5-7 days.
The invention also provides an application of the optimal factor obtained by the method for screening the optimal factor value in the human mesenchymal stem cell amplification method.
The invention has the beneficial effects that:
1. by the method, the optimal values of the amplification multiple and the cell viability rate are found;
2. by the method, a design space and a regression equation are obtained;
3. through the design space, the control parameter values which can approach the optimal amplification multiple and cell viability rate are obtained.
Drawings
FIG. 1 is a curved view of the amplification factor, cell seeding density and stirring speed of the present invention;
FIG. 2 is a contour plot of the amplification factor, cell seeding density and stirring speed in the present invention;
FIG. 3 is a curved surface diagram of cell viability, cell seeding density and stirring speed in the present invention;
FIG. 4 is a curved view of the cell viability, culture time and agitation speed in the present invention;
FIG. 5 is a contour plot of cell viability, culture time and agitation speed in the present invention.
Detailed Description
The present invention will be further described below, and it should be noted that the following examples are provided to illustrate the detailed embodiments and specific procedures based on the technical solution, but the scope of the present invention is not limited to the examples.
A screening method for optimal factor values in human mesenchymal stem cell expansion, the method comprising the steps of:
screening the S1 factor, and selecting the factor which possibly has a remarkable effect on the amplification of the human mesenchymal stem cells;
s2, carrying out single-factor variance analysis on the screened factors, and judging which factors are significance factors according to variance analysis results;
s3, carrying out variance analysis on the single factor in the step S2, and judging whether the single factor cannot be fitted into a straight line according to a regression equation; if not, returning to the step S1;
s4, if the fitting can be a straight line, performing a climbing experiment to find an interval where the optimal value is located;
s5, selecting 4 significant factors and intervals in the step S4, and carrying out RSM analysis;
s6, carrying out experimental design by using a BBD method, and obtaining data of amplification multiples and cell viability of all groups;
s7, performing ANOVA analysis on the data obtained in the step S6 to obtain a regression equation and a design space;
s8, respectively calculating the optimal factor values of the amplification multiple and the cell viability rate.
The factors screened in step S2 are 8, which are temperature, pH, dissolved oxygen, rotation speed, cell inoculation density, microcarrier inoculation density, culture time and medium replacement strategy.
It should be noted that the 4 significant factors and intervals in step S5 are the rotation speed, the cell seeding density, the microcarrier seeding density and the culture time, respectively.
The rotating speed is 20-40rpm, the cell inoculation density is 40000-120000cells/mL, the microcarrier inoculation density is 0.8-2.4g/L, and the culture time is 5-7 days.
The invention also provides an application of the optimal factor obtained by the method for screening the optimal factor value in the human mesenchymal stem cell amplification method.
Examples
The technical solution of the present invention will be further described by specific examples.
1. Single factor analysis
(1) And performing single-factor variance analysis on the 8 factors, and judging which factors are significance factors according to variance analysis results.
(2) And performing single-factor variance analysis on the 8 factors, judging whether the 8 factors cannot be fitted into a straight line according to a regression equation, and performing a climbing experiment to find an interval where the optimal value is located if the 8 factors can be fitted into the straight line.
2. RSM analysis
(1) And 4 significant factors and intervals are screened out to perform RSM analysis. Experimental design was performed using the BBD method, totaling 27 sets of experiments.
(2) 27 sets of experiments were performed, and 27 sets of data such as amplification factor, cell viability, etc. were obtained as shown in the following table:
(3) Carrying out ANOVA analysis on the 27 groups of data, analyzing the significance data of single factors, compound factors and square factors of the 27 groups of data, and removing insignificant items; if the residual data or fitting data is found to be unreasonable, the experimental group in question is re-experimented. Finally, a regression equation and a design space are obtained.
(4) And respectively calculating the optimal factor values of the amplification times and the cell viability rate. Wherein the amplification factor is calculated according to the following formula:
amplification multiple = -14.71+0.348 stirring rotation speed +0.000124 cell inoculation density +3.28 microcarrier inoculation density +1.82 culture time-0.00267 stirring rotation speed-0.000000 cell inoculation density: -1.413 microcarrier inoculation density: -0.104 culture time: +0.000000 stirring rotation speed: -0.0219 stirring rotation speed: -0.0250 stirring rotation speed: -0.000001 cell inoculation density: +0.344 microcarrier inoculation density.
Cell viability = -0.085+0.0159 stirring rotation speed +0.000003 cell inoculation density +0.132 microcarrier inoculation density +0.212 culture time-0.000275 stirring rotation speed-0.000000 cell inoculation density-0.0137 microcarrier inoculation density-0.0162 culture time +0.000000 stirring rotation speed +cell inoculation density +0.00031 stirring rotation speed:microcarrierinoculation density-0.00100 stirring rotation speed: -0.000001 cell inoculation density +0.000000 cell inoculation density-0.0031 microcarrier inoculation density.
Simulation test
And (3) carrying out verification experiments according to the optimal factor value designed by the technical scheme, wherein if the optimal factor value is close to the optimal values of the amplification multiple and the cell viability, the experiments are successful.
If not, analyzing the experimental process, performing problem troubleshooting, and performing the experiment again until the optimal value is approached.
Various corresponding changes and modifications can be made by those skilled in the art based on the above technical solutions and concepts, and all such changes and modifications should be included in the protection scope of the present invention.
Claims (5)
1. A screening method for optimal factor values in human mesenchymal stem cell expansion, characterized in that the method comprises the following steps:
screening S1 factors, and selecting factors which can have obvious effect on the amplification of the human mesenchymal stem cells;
s2, carrying out single-factor variance analysis on the screened factors, and judging which factors are significant factors according to variance analysis results;
s3, carrying out variance analysis on the single factor in the step S2, and judging whether the single factor cannot be fitted into a straight line according to a regression equation; if not, returning to the step S1;
s4, if the fitting can be a straight line, performing a climbing experiment to find an interval where the optimal value is located;
s5, selecting 4 significant factors and intervals in the step S4, and carrying out RSM analysis;
s6, carrying out experimental design by using a BBD method, and obtaining data of amplification multiples and cell viability rates of all groups;
s7, performing ANOVA analysis on the data obtained in the step S6 to obtain a regression equation and a design space;
s8, respectively calculating the optimal factor values of the amplification multiple and the cell viability rate.
2. The method for screening optimal factor values for human mesenchymal stem cell expansion according to claim 1, wherein the number of the factors screened in step S2 is 8, and the factors are temperature, pH, dissolved oxygen, rotation speed, cell inoculation density, microcarrier inoculation density, culture time and culture medium fluid replacement strategy.
3. The screening method for optimal factor values in human mesenchymal stem cell expansion according to claim 1, wherein the 4 significant factors and intervals in step S5 are rotation speed, cell seeding density, microcarrier seeding density and culture time, respectively.
4. The screening method for optimal factor value in human mesenchymal stem cell expansion according to claim 3, wherein the rotation speed is 20-40rpm, the cell seeding density is 40000-120000cells/mL, the microcarrier seeding density is 0.8-2.4g/L, and the culture time is 5-7 days.
5. The optimal factor obtained by the screening method of claim 1 is applied to a human mesenchymal stem cell amplification method.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103361308A (en) * | 2012-03-30 | 2013-10-23 | 华东理工大学 | Method for amplifying mesenchymal stem cells on large scale |
| CN105001009A (en) * | 2015-08-13 | 2015-10-28 | 福建农林大学 | Antrodia cinnamomea terpene-rich mycelium culture medium and application |
| CN105543313A (en) * | 2015-12-29 | 2016-05-04 | 四川新生命干细胞科技股份有限公司 | Human-derived mesenchymal stem cell factor, and preparation method and application thereof |
| CN105838627A (en) * | 2016-05-10 | 2016-08-10 | 湖北省烟草公司恩施州公司 | Penicillium xfsf-8 strain for preventing and treating tobacco bacterial wilt and application |
| US20180280441A1 (en) * | 2015-04-03 | 2018-10-04 | Stemmedicare Co., Ltd. | Method for mass producing proteins in mesenchymal stem cells |
| CN110093312A (en) * | 2019-05-15 | 2019-08-06 | 张永国 | A kind of cell large-scaled culture method, purification process and cell messenger |
| CN110804587A (en) * | 2019-11-27 | 2020-02-18 | 浙江卫未生物医药科技有限公司 | Method for modeling production of mesenchymal stem cells by adopting improved microcarrier cell culture method |
| CN110923203A (en) * | 2019-12-03 | 2020-03-27 | 北京银丰鼎诚生物工程技术有限公司 | Amplification culture process of umbilical cord mesenchymal stem cells |
| CN113755515A (en) * | 2021-09-29 | 2021-12-07 | 江南大学 | Recombinant escherichia coli for efficiently producing clavulanic acid and application thereof |
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- 2022-10-21 CN CN202211294728.0A patent/CN115466724A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103361308A (en) * | 2012-03-30 | 2013-10-23 | 华东理工大学 | Method for amplifying mesenchymal stem cells on large scale |
| US20180280441A1 (en) * | 2015-04-03 | 2018-10-04 | Stemmedicare Co., Ltd. | Method for mass producing proteins in mesenchymal stem cells |
| CN105001009A (en) * | 2015-08-13 | 2015-10-28 | 福建农林大学 | Antrodia cinnamomea terpene-rich mycelium culture medium and application |
| CN105543313A (en) * | 2015-12-29 | 2016-05-04 | 四川新生命干细胞科技股份有限公司 | Human-derived mesenchymal stem cell factor, and preparation method and application thereof |
| CN105838627A (en) * | 2016-05-10 | 2016-08-10 | 湖北省烟草公司恩施州公司 | Penicillium xfsf-8 strain for preventing and treating tobacco bacterial wilt and application |
| CN110093312A (en) * | 2019-05-15 | 2019-08-06 | 张永国 | A kind of cell large-scaled culture method, purification process and cell messenger |
| CN110804587A (en) * | 2019-11-27 | 2020-02-18 | 浙江卫未生物医药科技有限公司 | Method for modeling production of mesenchymal stem cells by adopting improved microcarrier cell culture method |
| CN110923203A (en) * | 2019-12-03 | 2020-03-27 | 北京银丰鼎诚生物工程技术有限公司 | Amplification culture process of umbilical cord mesenchymal stem cells |
| CN113755515A (en) * | 2021-09-29 | 2021-12-07 | 江南大学 | Recombinant escherichia coli for efficiently producing clavulanic acid and application thereof |
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