CN113151158A - Tissue stem cell separation and function evaluation system - Google Patents
Tissue stem cell separation and function evaluation system Download PDFInfo
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- 238000000926 separation method Methods 0.000 title claims abstract description 23
- 238000011156 evaluation Methods 0.000 title claims abstract description 21
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- 210000000287 oocyte Anatomy 0.000 claims abstract description 49
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Abstract
The invention discloses a tissue stem cell separation and function evaluation system, which comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst; after the blastocyst is cultured, the zona pellucida or part thereof is removed from the blastocyst, then the trophoblast is removed from the obtained blastocyst to separate the inner cell mass, the gene sequence of the white mouse is similar to that of human, the whole genome of the white mouse has extremely high similarity with human, and many diseases which are difficult to cure by human can find similar characters on the white mouse, so that the experiment is carried out to find the disease curing gene, and the functions of the separated components of the tissue stem cell can be obtained more easily.
Description
Technical Field
The invention belongs to the technical field of stem cell separation, and particularly relates to a tissue stem cell separation and function evaluation system.
Background
Stem cells are a type of pluripotent cells with self-replication ability, which can be differentiated into various functional cells under certain conditions, and are divided into embryonic stem cells and adult stem cells according to the development stage of the stem cells, and can be divided into three types according to the development potential of the stem cells: the stem cell is an insufficiently differentiated and immature cell, has potential functions of regenerating various organs and human bodies, and is called as a universal cell in the medical field.
In the prior art, manual operation is mostly adopted during cell stem cell separation, the dosage of normal saline and stem cell stock solution is difficult to accurately control, manual operation causes that the operating environment is generally uncontrolled, the separating medium that can influence the separation is not pure enough like this, adopt manual operation, lead to separation inefficiency, and make mistakes easily, operating personnel adopts the artifical separating medium that absorbs the different degree of depth one by one to the centrifugation of tissue stem cell to draw, this kind of mode can not ensure that the supernatant draws totally, if the straw inserts deeply, then can inhale lower floor's precipitate together, the purity of difficult control separating medium, cell separation efficiency is lower, for this reason, we propose a tissue stem cell separation and functional evaluation system.
Disclosure of Invention
The technical problem to be solved by the present invention is to overcome the existing defects, and provide a tissue stem cell separation and function evaluation system, so as to solve the problems that in the prior art, manual operation is mostly adopted during cell stem cell separation, the usage amount of physiological saline and stem cell stock solution is difficult to accurately control, and the manual operation causes the general uncontrolled operation environment, which affects that the separated separation solution is not pure enough, and the manual operation causes low separation efficiency and is easy to make mistakes.
In order to achieve the purpose, the invention provides the following technical scheme: a tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
Preferably, the mouse in the first step is an adult mouse, the oocyte is extracted 5 days after mating, and the embryo is cultured in a culture dish filled with a culture solution.
Preferably, the concentration of calcium ions used in the second step is 10. mu.M, and the concentration of 6-dimethylaminopurine used in the second step is 2. mu.M.
Preferably, in the sixth step, the tissue stem cells are amplified by a PCR amplification machine, and then the purified small molecule compound is subjected to T-cloning treatment.
Preferably, the T-vector is an overhang vector, and the PCR product can be rapidly and directly inserted into the multiple cloning site of the plasmid vector in one step under the action of enzyme.
Preferably, the T vector is a special plasmid vector for efficiently cloning a PCR product, is a linearized vector, and can be directly connected with the PCR product without enzyme digestion to obtain a corresponding small molecular compound.
Compared with the prior art, the invention provides a tissue stem cell separation and function evaluation system, which has the following beneficial effects:
1. the present invention relates to a method for producing a mouse, which comprises the steps of mating a male mouse and a female mouse, allowing the fertilized female mouse to stand for several days, obtaining an oocyte in the fertilized mouse, reprogramming, activating and culturing the obtained oocyte in vitro to form a blastocyst, activating the oocyte by using a calcium ionophore having a concentration of 5 μ M to 15 μ M, and then treating the oocyte by using 6-dimethylaminopurine having a concentration of 1 μ M to 5 μ M, performing in vitro culture by using two different media, namely a G1.2 medium and a SNUnt-2 medium, removing a zona pellucida or a portion thereof from the blastocyst after completion of the culture, removing a feeder layer from the obtained blastocyst to isolate an inner cell mass, wherein the gene sequence of the mouse is almost the same as that of human, the whole genome thereof has a very high similarity to human, and many diseases which are difficult to cure by human can find similar traits in the mouse, experiments are carried out to find disease treatment genes, so that the functions of the separated components of the tissue stem cells can be obtained more easily;
2. according to the invention, small molecular compounds obtained by amplifying specific tissue stem cells are researched and recorded, the functional characteristics of the small molecular compounds are determined, then the capability of forming microstructures of the small molecular compounds is evaluated by referring to pluripotent stem cells, different biological models are established, the stem cells are transplanted into the biological models, the functions of the stem cells entering the different models are preliminarily determined by observing and detecting the models, the functions are evaluated functionally, and finally more than two new schemes for treating major diseases by transplanting the tissue stem cells and the structures formed by the tissue stem cells are established.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention without limiting the invention in which:
FIG. 1 is a flow chart of a tissue stem cell isolation and function evaluation system according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a technical scheme that: a tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
And (3) taking the oocytes 5 days after mating of the mice in the first step, and culturing the taken embryos in a culture dish filled with a culture solution.
The concentration of calcium ions used in step two was 10. mu.M, and the concentration of 6-dimethylaminopurine used in step two was 2. mu.M.
And in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
The T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
The T vector is a special plasmid vector for efficiently cloning PCR products, is a linearized vector, and can be directly connected with the PCR products without enzyme digestion to obtain corresponding small molecular compounds.
Example one
A tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
And (3) taking the oocytes 5 days after mating of the mice in the first step, and culturing the taken embryos in a culture dish filled with a culture solution.
The concentration of calcium ions used in step two was 10. mu.M, and the concentration of 6-dimethylaminopurine used in step two was 2. mu.M.
And in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
The T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
The T vector is a special plasmid vector for efficiently cloning PCR products, is a linearized vector, and can be directly connected with the PCR products without enzyme digestion to obtain corresponding small molecular compounds.
EXAMPLE III
A tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
And (3) taking the oocytes 5 days after mating of the mice in the first step, and culturing the taken embryos in a culture dish filled with a culture solution.
The concentration of calcium ions used in step two was 10. mu.M, and the concentration of 6-dimethylaminopurine used in step two was 2. mu.M.
And in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
The T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
The T vector is a special plasmid vector for efficiently cloning PCR products, is a linearized vector, and can be directly connected with the PCR products without enzyme digestion to obtain corresponding small molecular compounds.
Example four
A tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
And (3) taking the oocytes 5 days after mating of the mice in the first step, and culturing the taken embryos in a culture dish filled with a culture solution.
The concentration of calcium ions used in step two was 10. mu.M, and the concentration of 6-dimethylaminopurine used in step two was 2. mu.M.
And in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
The T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
The T vector is a special plasmid vector for efficiently cloning PCR products, is a linearized vector, and can be directly connected with the PCR products without enzyme digestion to obtain corresponding small molecular compounds.
EXAMPLE five
A tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
And (3) taking the oocytes 5 days after mating of the mice in the first step, and culturing the taken embryos in a culture dish filled with a culture solution.
The concentration of calcium ions used in step two was 10. mu.M, and the concentration of 6-dimethylaminopurine used in step two was 2. mu.M.
And in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
The T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
The T vector is a special plasmid vector for efficiently cloning PCR products, is a linearized vector, and can be directly connected with the PCR products without enzyme digestion to obtain corresponding small molecular compounds.
EXAMPLE six
A tissue stem cell isolation and functional assessment system comprising the steps of:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
And (3) taking the oocytes 5 days after mating of the mice in the first step, and culturing the taken embryos in a culture dish filled with a culture solution.
The concentration of calcium ions used in step two was 10. mu.M, and the concentration of 6-dimethylaminopurine used in step two was 2. mu.M.
And in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
The T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
The T vector is a special plasmid vector for efficiently cloning PCR products, is a linearized vector, and can be directly connected with the PCR products without enzyme digestion to obtain corresponding small molecular compounds.
The working principle and the using process of the invention are as follows: in use, a male mouse and a female mouse are placed together to mate the male mouse and the female mouse, the fertilized female mouse is kept still for several days to obtain an oocyte in the fertilized mouse, the obtained oocyte is reprogrammed, activated and cultured in vitro to form a blastocyst, the oocyte is activated by using a calcium ion carrier with the concentration of 10 mu M and then treated by using 6-dimethylaminopurine with the concentration of 2 mu M, the in vitro culture is carried out by using two different culture media, namely a G1.2 culture medium and a SNUnt-2, after the culture is finished, a zona pellucida or a part thereof is removed from the blastocyst, an inner cell mass is separated from the obtained blastocyst by removing a trophoblast, the obtained blastocyst is placed into a device for centrifugal treatment, stem cell separating solutions with different depths obtained by centrifugation are respectively placed into different culture dishes for culture, and stem cells with specific tissues are detected and screened, researching and recording different functional characteristics of the specific tissue stem cells, and amplifying effective small molecular compounds by using different specific tissue stem cells, and the functional characteristics of the small molecular compounds are researched and recorded, and after the characteristics of the small molecular compounds are determined, evaluating the capability of forming microstructures of the pluripotent stem cells by referring to the pluripotent stem cells, establishing different biological models, transplanting the stem cells into the biological models, through observation and detection of the models, the functions of stem cells entering different models are preliminarily determined, and performing functional assessment, re-establishing different disease biological models, transplanting stem cells into the models, observing and determining the scheme and the result of the stem cells for treating diseases, and finally establishing more than two new schemes for treating major diseases by tissue stem cells and forming structure transplantation.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. A tissue stem cell isolation and functional assessment system, comprising: the method comprises the following steps:
the method comprises the following steps: mating male and female mice, resting the fertilized female mouse for several days, taking an oocyte from the fertilized mouse, reprogramming, activating and culturing the oocyte in vitro to form a blastocyst;
step two: activating the oocyte by using calcium ionophore at a concentration of 5 μ M to 15 μ M and subsequently treating the oocyte with 6-dimethylaminopurine at a concentration of 1 μ M to 5 μ M;
step three: performing in vitro culture by using two different culture media, namely G1.2 culture medium and SNUnt-2 culture medium, removing a zona pellucida or a part thereof from a blastocyst, and removing a feeder layer from the resulting blastocyst to isolate an inner cell mass;
step four: placing the obtained blastocyst into a centrifugal device for centrifugal treatment, and placing stem cell separation solutions with different depths obtained by centrifugation into different culture dishes for culture;
step five: detecting and screening stem cells with specific tissues, and researching and recording different functional characteristics of the stem cells with the specific tissues;
step six: then different specific tissue stem cells are utilized to amplify effective small molecular compounds, the functional characteristics of the small molecular compounds are researched and recorded, and the capability of forming a microstructure is evaluated by referring to the pluripotent stem cells after the characteristics of the small molecular compounds are determined;
step seven: establishing different biological models, transplanting the stem cells into the biological models, preliminarily determining the functions of the stem cells entering the different models through observation and detection of the models, and performing functional evaluation;
step eight: establishing different disease biological models again, transplanting the stem cells into the models, and observing and determining the scheme and the result of the stem cells for treating the diseases;
step nine: finally, more than two new schemes for treating serious diseases through tissue stem cells and the transplantation of the tissue stem cells forming structures are established.
2. The tissue stem cell isolation and function evaluation system of claim 1, wherein: the mice in the first step are newly-grown mice, oocytes are extracted 5 days after mating, and the extracted embryos are placed in a culture dish filled with a culture solution for culture.
3. The tissue stem cell isolation and function evaluation system of claim 1, wherein: the concentration of calcium ions used in the second step was 10. mu.M, and the concentration of 6-dimethylaminopurine used in the second step was 2. mu.M.
4. The tissue stem cell isolation and function evaluation system of claim 1, wherein: and in the sixth step, the tissue stem cells are amplified by a PCR (polymerase chain reaction) amplification instrument, and then the purified small molecular compounds are subjected to T-cloning treatment.
5. The tissue stem cell isolation and function evaluation system of claim 3, wherein: the T vector is a vector with an overhang, and can rapidly insert the PCR product into the multiple cloning sites of the plasmid vector in one step under the action of enzyme.
6. The tissue stem cell isolation and function evaluation system of claim 3, wherein: the T vector is a special plasmid vector for efficiently cloning a PCR product, is a linearized vector, and can be directly connected with the PCR product without enzyme digestion to obtain a corresponding small molecular compound.
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