CN109529054A - The evaluation method and application of mescenchymal stem cell preparation oncogenicity - Google Patents
The evaluation method and application of mescenchymal stem cell preparation oncogenicity Download PDFInfo
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Abstract
The present invention provides the evaluation methods and application of a kind of mescenchymal stem cell preparation oncogenicity, are related to field of biotechnology.The evaluation method of the mescenchymal stem cell preparation includes providing the animal subject for receiving the mescenchymal stem cell preparation administration, and the oncogenicity of mescenchymal stem cell is then evaluated according to pre-set level；Wherein, the dosage for receiving the animal subject of mescenchymal stem cell preparation administration is 0.5 × 106~5 × 106Cells/ is only；Pre-set level includes following index: (a1) changes of weight；(a2) organ weights and organ coefficient；(a3) whether nodosity is formed in the animal subject body.The evaluation method alleviate it is existing in the prior art lack it is a kind of effectively to the evaluation method of the oncogenicity of mescenchymal stem cell preparation the problem of.
Description
Technical field
The present invention relates to field of biotechnology, more particularly, to a kind of evaluation method of mescenchymal stem cell preparation oncogenicity
And application.
Background technique
Mescenchymal stem cell is a kind of multipotential stem cell, from the mesoderm and ectoderm of mesoderm growing early stage, has self
It updates and Multidirectional Differentiation ability, the interstitial tissues such as promotion bone, cartilage, ligament, muscle and adipose tissue regenerates.Mescenchymal stem cell
Also there is its special biological property, comprising: immunoregulation effect；Point of hemopoiesis-supporting founction and various bioactivators
Secrete ability.Mescenchymal stem cell, which is referred to as, enters clinical test, treats immunity disease and Various Tissues damage related disease, and
It most promises to be after candidate stem cell, into another adult stem cell in clinical application stage.Because of mescenchymal stem cell
Abundance prepares simple, versatility and low oncogenicity with very big clinical utility.Currently, being filled between many countries
Matter stem cell has been approved by into multiple clinical trials, for treating graft versus host disease(GVH disease), Crohn's disease, hepatic fibrosis-renal tubular ectasia syndrome
With a variety of diseases such as osteoarticular injury.It is external easily operated since source for mesenchymal stem cells is extensive, hematopoiesis support and immune tune
Section effect is clear, therefore mescenchymal stem cell treatment has broad prospects.
Although mescenchymal stem cell comes into III clinical trial phase, safety issue still needs to be paid attention to.Mesenchyma
Evaluation of the stem cell in terms of preclinical safety evaluatio, especially oncogenicity lacks evaluation index and corresponding technology hand
Section.Therefore a kind of evaluation method of effective mescenchymal stem cell preparation oncogenicity be guarantee mescenchymal stem cell related drugs and
The guarantee for the treatment of method clinically security application.
In view of this, the present invention is specifically proposed.
Summary of the invention
The first object of the present invention is to provide a kind of evaluation method of mescenchymal stem cell preparation oncogenicity, alleviate existing
Have and lacks the technical issues of oncogenicity evaluation effectively is carried out to mescenchymal stem cell preparation present in technology.
The second object of the present invention is the evaluation method for providing mescenchymal stem cell preparation oncogenicity between preparation includes
Application in the drug of mesenchymal stem cells.
In order to solve the above technical problems, spy of the present invention adopts the following technical scheme that
The present invention provides a kind of evaluation method of mescenchymal stem cell preparation oncogenicity, the evaluation method includes: to mention
For receiving the animal subject of the mescenchymal stem cell preparation administration, the cause of mescenchymal stem cell is then evaluated according to pre-set level
Tumor；
The dosage of the animal subject for receiving the mescenchymal stem cell preparation administration is 0.5 × 106~5 ×
106Cells/ is only；
The pre-set level includes following index:
(a1) changes of weight；
(a2) organ weights and organ coefficient；
(a3) whether nodosity is formed in the animal subject body.
Preferably, it the time for receiving the mescenchymal stem cell preparation administration with the animal subject, obtains weekly
The weight of the animal subject.
Preferably, it after starting 10 weeks with the time that the animal subject receives the mescenchymal stem cell preparation administration, obtains
Take the organ weights and organ coefficient of the animal subject；
With, after starting 6 months with the time that the animal subject receives the mescenchymal stem cell preparation administration, acquisition institute
State the organ weights and organ coefficient of animal subject；
Preferably, the internal organs include one or more of the heart, liver, spleen, lung, kidney and thymus gland.
Preferably, after starting 10 weeks with the time that the animal subject receives the mescenchymal stem cell preparation administration, inspection
It looks into the animal subject body and whether tubercle occurs；
With, after starting 6 months with the time that the animal subject receives the mescenchymal stem cell preparation administration, inspection institute
It states in animal subject body and whether tubercle occurs；
Preferably, described to check that whether occurring tubercle in the animal subject body includes following (b1) and/or (b2):
(b1) it is dissected after putting to death animal subject, whether nodosity is formed observation internal organs；
(b2) the progress Histopathology detection of animal subject internal organs is obtained after putting to death animal subject.
Preferably, the evaluation method further includes blank control group；The blank control group include receive physiological saline to
The animal subject of medicine；
Preferably, the blank control group and the animal subject difference for receiving the mescenchymal stem cell preparation administration
Independent includes 3~5 animal subjects.
Preferably, the animal subject includes mouse, rat, cavy, rabbit, dog, miniature pig, monkey, ferret, marmot and naked
One or more of mole；Preferably mouse.
Preferably, the animal subject is BALB/c Nude mouse；The BALB/c Nude mouse is 6-8 week old, weight
Amount is the female mice of 18~20g.
Preferably, the dosage of the BALB/c Nude mouse is 1 × 106~2.5 × 106Cells/ is only；Preferably 2
×106Cells/ is only.
Preferably, the administration mode of the administration include subcutaneous administration, intramuscular administration, intravenous administration, intravaginal administration,
Intrauterine administration, pulmonary administration or rectally；
Preferably, the administration mode includes intravenous administration；More preferably mouse tail vein input administration.
It is dry comprising mesenchyma in preparation that the present invention also provides the evaluation methods of above-mentioned mescenchymal stem cell preparation oncogenicity
Application in the drug of cell；
Preferably, the drug includes human umbilical cord mesenchymal stem cells.
Compared with prior art, the invention has the following beneficial effects:
The evaluation method of mescenchymal stem cell preparation oncogenicity provided by the invention, on the one hand provide one reasonably to
The dosage range of medicine avoids the problem due to the unreasonable caused evaluation result inaccuracy of dosage；On the other hand, this method
Changes of weight of the overall merit animal subject after receiving the administration of mescenchymal stem cell preparation, organ weights and organ coefficient
And whether nodosity forms totally three Xiang Zhibiao in animal subject body, keeps the setting of evaluation index more scientific and reasonable.Traditional cause
Tumor evaluation often only evaluates in animal subject body whether form tubercle, and has ignored animal subject during the experiment due to it
The problem of physical function caused by his factor declines, and animal subject is made more to be susceptible to suffer from tumour.The application overall merit animal subject is given
Body variation after medicine, is not concerned only with whether animal subject tubercle occurs in vivo, and the sign of also concern animal subject changes, comprehensive
That closes has rated mescenchymal stem cell preparation to the tumorigenesis degree of animal subject.Conceived based on foregoing invention, the present invention also provides
Application of the evaluation method of mescenchymal stem cell preparation oncogenicity in drug of the preparation comprising mescenchymal stem cell, to assist
Exploitation of the mescenchymal stem cell as active constituent or the drug of auxiliary element.
Detailed description of the invention
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art be briefly described, it should be apparent that, it is described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, is also possible to obtain other drawings based on these drawings.
Figure 1A is Normal group provided in an embodiment of the present invention and the weight of mouse becomes after the administration of MSC injection group 10 weeks
Change；
Figure 1B is the weight that the mouse in 6 months is administered in Normal group provided in an embodiment of the present invention and MSC injection group
Variation；
Fig. 2A is the observation after Normal group provided in an embodiment of the present invention is administered 10 weeks to mice organs, nothing in internal organs
Macroscopic tubercle is formed；
Fig. 2 B is the observation after MSC injection group provided in an embodiment of the present invention is administered 10 weeks to mice organs, nothing in internal organs
Macroscopic tubercle is formed；
Fig. 2 C is observation to mice organs after Normal group provided in an embodiment of the present invention is administered 6 months, in internal organs
The tubercle being visible by naked eyes is formed；
Fig. 2 D is the observation after MSC injection group provided in an embodiment of the present invention is administered 6 months to mice organs, nothing in internal organs
Macroscopic tubercle is formed；
The internal organs system of Fig. 3 A mouse core for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 10 weeks
Number；
The internal organs system of Fig. 3 B Mouse Liver for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 10 weeks
Number；
The internal organs system of Fig. 3 C mice spleen for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 10 weeks
Number；
The internal organs system of Fig. 3 D mouse lung for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 10 weeks
Number；
The internal organs system of Fig. 3 E Mouse Kidney for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 10 weeks
Number；
The internal organs of Fig. 4 A mouse core for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 6 months
Coefficient；
The internal organs of Fig. 4 B Mouse Liver for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 6 months
Coefficient；
The internal organs of Fig. 4 C mice spleen for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 6 months
Coefficient；
The internal organs of Fig. 4 D mouse lung for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 6 months
Coefficient；
The internal organs of Fig. 4 E Mouse Kidney for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 6 months
Coefficient；
Fig. 5 A is for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 10 weeks, mouse main organs
Histopathology detection；
Fig. 5 B is for Normal group provided in an embodiment of the present invention and after the administration of MSC injection group 6 months, mouse main organs
Histopathology detection.
Specific embodiment
Technical solution of the present invention is clearly and completely described below in conjunction with attached drawing, it is clear that described implementation
Example is a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, ordinary skill
Personnel's every other embodiment obtained without making creative work, shall fall within the protection scope of the present invention.
The person that is not specified actual conditions in embodiment, carries out according to conventional conditions or manufacturer's recommended conditions.Agents useful for same or instrument are not
Production firm person is indicated, is the conventional products that can be obtained by commercially available purchase.
In the present invention, if without particularly illustrating, all embodiments mentioned in this article and preferred implementation method
It can be combined with each other to form new technical solution.
In the present invention, if without particularly illustrating, all technical characteristics and preferred feature mentioned in this article can be with
Intercombination forms new technical solution.
In the present invention, if percentage (%) or part refer to the weight relative to composition without particularly illustrating
Percentage or parts by weight.
In the present invention, if related each component or its preferred ingredient can be combined with each other shape without particularly illustrating
The technical solution of Cheng Xin.
In the present invention, unless otherwise indicated, numberical range " a~b " indicates the contracting of any real combinings between a to b
Sketch form shows that wherein a and b is real number.Such as numberical range " 6~22 " indicate herein all listed " 6~22 " it
Between whole real numbers, " 6~22 " be these combinations of values breviary indicate.
" range " disclosed in this invention can be respectively one or more lower limits and one in the form of lower and upper limit
A or multiple upper limits.
In the present invention, unless otherwise indicated, it is each reaction or operating procedure can sequentially carry out, can also in sequence into
Row.Preferably, reaction method herein is that sequence carries out.
The present invention provides a kind of evaluation method of mescenchymal stem cell preparation oncogenicity, the evaluation method includes: to mention
For receiving the animal subject of the mescenchymal stem cell preparation administration, the cause of mescenchymal stem cell is then evaluated according to pre-set level
Tumor；
The dosage of the animal subject for receiving the mescenchymal stem cell preparation administration is 0.5 × 106~5 ×
106Cells/ only, such as can be but be not limited to 0.5 × 106Cells/, 1 × 106Cells/, 1.5 × 106cells/
Only, 2 × 106Cells/, 2.5 × 106Cells/, 3 × 106Cells/, 3.5 × 106Cells/, 4 × 106cells/
Only, 4.5 × 106Cells/, 5 × 106Cells/, 5.5 × 106Cells/ or 6 × 106Cells/ is only；
The pre-set level includes following index: (a1) changes of weight；(a2) organ weights and organ coefficient；(a3) institute
State in animal subject body whether nodosity is formed.
Mescenchymal stem cell is a kind of multipotential stem cell, have self-renewing and Multidirectional Differentiation ability, promote bone, cartilage,
The regeneration of the interstitial tissues such as ligament, muscle and adipose tissue.Because source for mesenchymal stem cells is abundant, preparation is simple and versatility has
Very big clinical utility.The evaluation method of mescenchymal stem cell preparation oncogenicity provided by the invention, on the one hand provides
One dosage range being reasonably administered avoids the problem due to the unreasonable caused evaluation result inaccuracy of dosage；Separately
On the one hand, changes of weight of this method overall merit animal subject after receiving the mescenchymal stem cell preparation administration,
Whether nodosity forms totally three Xiang Zhibiao in organ weights and organ coefficient and the animal subject body, this three indexs can be complete
Mescenchymal stem cell is evaluated in face, and whether to mouse body, there are adverse effects, if to each organ, there are adverse effects, if
There are oncogenic functions in each organ, keep the setting of evaluation index more scientific and reasonable.Traditional oncogenicity evaluation is often only commented
Tubercle whether is formed in valence animal subject body, and has ignored the animal subject body as caused by other factors during the experiment
The problem of function declines, and animal subject is made more to be susceptible to suffer from tumour.Body variation after the administration of the application overall merit animal subject, no
It is concerned only with whether animal subject tubercle occurs in vivo, the sign of also concern animal subject changes, and comprehensive has rated mesenchyma
Tumorigenesis degree of the stem cell medicine to animal subject.
It should be noted that mescenchymal stem cell preparation of the present invention refers to the examination comprising human mesenchymal stem cell
Agent, drug or mescenchymal stem cell itself；Dosage of the present invention is to play work in mescenchymal stem cell preparation
Property ingredient mescenchymal stem cell quantity calculate.The mescenchymal stem cell, which is isolated from, for example can be but be not limited to bone
Marrow, periosteum, blood vessel, fat, muscle, peripheral circulation, Cord blood, skin or tissue of tooth etc.；The mescenchymal stem cell is for example
It can be but be not limited to the isolated cell in laboratory or commercially available cell line；The source of the mescenchymal stem cell is for example
Can be but be not limited to for people or other separate the animal subject of mescenchymal stem cell；The mescenchymal stem cell is for example
Can be but be not limited to obtain by the acceptable means in this field through mutagenesis, mutation or being transformed through genetic engineering
The mescenchymal stem cell arrived.It is understood that the purpose of the present invention is evaluating the oncogenicity of mescenchymal stem cell preparation, therefore
With no restrictions to the source of mescenchymal stem cell.
In some preferred embodiments, with the animal subject receive mescenchymal stem cell preparation administration when
Between start, obtain the weight of the animal subject weekly.
In some preferred embodiments, with the animal subject receive mescenchymal stem cell preparation administration when
Between start 10 weeks after, obtain the organ weights and organ coefficient of the animal subject；With, with the animal subject receive it is described between
After the time of mesenchymal stem cells preparation administration starts 6 months, the organ weights and organ coefficient of the animal subject are obtained；Its
In, organ coefficient=organ weight/weight；The internal organs preferably include the Other Main Internal Organs of animal subject, such as can be
One or more of but be not limited to the heart, liver, spleen, lung, kidney and thymus gland.
In some preferred embodiments, with the animal subject receive mescenchymal stem cell preparation administration when
Between start 10 weeks after, check in the animal subject body whether tubercle occur；With the mesenchyma is received with the animal subject
After the time of stem cell medicine administration starts 6 months, check in the animal subject body whether tubercle occur.It is preferred that according to as follows
At least one of two methods check in the animal subject body whether tubercle occur: (b1) is dissected after putting to death animal subject,
Observing internal organs, whether nodosity is formed, and dissection can directly observe the viscera of animal subject；(b2) it is obtained after putting to death mouse
Mice organs are taken to carry out Histopathology detection, to further determine that the disease of animal subject.
In some preferred embodiments, the evaluation method further includes that setting blank control group progress methodology is tested
Card, to further decrease error of quality appraisement；The blank control group includes the animal subject for receiving saline administration.Some
In preferred embodiment, the blank control group and the animal subject point for receiving the mescenchymal stem cell preparation administration
Not independent includes 3~5 animal subjects.
In some preferred embodiments, the animal subject include mouse, rat, cavy, rabbit, dog, miniature pig,
One or more of monkey, ferret, marmot and naked mole；Preferably mouse.The animal subject is preferably BALB/c
Nude mouse；The BALB/c Nude mouse is 6-8 week old, and weight is the female mice of 18~20g.
In some preferred embodiments, the BALB/c Nude for receiving the mescenchymal stem cell preparation administration
The dosage of mouse is 1 × 106~2.5 × 106Cells/ only, such as can be but be not limited to 1 × 106Cells/,
1.5×106Cells/, 2 × 106Cells/ or 2.5 × 106Cells/ is only；Preferred dosage is 2 × 106cells/
Only.
In some preferred embodiments, the administration mode of the administration include subcutaneous administration, it is intramuscular administration, intravenous
Administration, intravaginal administration, intrauterine administration, pulmonary administration or rectally；Preferably, the administration mode includes intravenously giving
Medicine；More preferably mouse tail vein input administration.
It is dry comprising mesenchyma in preparation that the present invention also provides the evaluation methods of above-mentioned mescenchymal stem cell preparation oncogenicity
Application in the drug of cell, to assist the exploitation using mescenchymal stem cell as active constituent or the drug of auxiliary element.
In some preferred embodiments, the drug includes human umbilical cord mesenchymal stem cells, human umbilical cord mesenchymal stem cells
(human umbilical cord mesenchymal stem cells, hUC-MSCs) is to be present in neonatal umbilical cord tissue
One of versatile stem cell, be a kind of low immunogenicity cell, and have very strong immunoloregulation function.Umbilical cord mesenchyma
Stem cell possesses the complete characteristic of MSC, and rich content, is easily isolated culture.Compared with the MSC in other sources, UC-MSC
It is more original, it is the stem cell between embryonic stem cell and adult stem cell, proliferation and differentiation capability compare embryonic stem cell
It is low, but it is apparently higher than adult stem cell.Umbilical cord mesenchymal stem cells have more differentiation potentials, under specific inductive condition, UC-
MSC can not only be divided into the mesoblastemas such as bone, cartilage, fat, tendon, and being capable of inside embryonic tissue cell (such as cardiac muscle
Cell, liver cell) and ectoderm tissue cell (such as nerve cell) differentiation, it can be in different inductive conditions and suitable life in vivo
In long microenvironment, safely directed differentiation is different tissue lines, has the ability for repairing various tissues and organ.Therefore
In the application for preparing drug all the more extensively, the evaluation method using above-mentioned mescenchymal stem cell preparation oncogenicity can be UC-MSC
UC-MSC provides safety data in the research for preparing drug, is reference data for clinical drug use.
Beneficial effects of the present invention are further illustrated below with reference to preferred embodiment.
Embodiment
The mescenchymal stem cell preparation of the present embodiment evaluation is human umbilical cord mesenchymal stem cells (MSC), that is, is sent to and is administered.
Experimental design:
(1) animal subject: BALB/c Nude mouse, 6-8 week old, female, 19 ± 1g are tested.
(2) group is arranged: whole mouse are randomly divided into Normal group (control) and MSC injection group, Normal group
Each 4 with MSC injection group.
(3) administration route: tail vein administration.
(4) dosage: 200 μ l sodium chloride injection of Normal group (control) tail vein injection；MSC injection group
Tail vein injection 2 × 106It is a/only MSC.
Test method:
(1) animal subject is tested:
Animal subject receives: experiment animal subject ensures that departmental staff and animal doctor connect jointly by experimenter, animal subject
It receives.Means of transport meets the requirements when reception, and the animal subject verification of conformity content and application that animal subject supplying unit provides are purchased
Animal subject kind, rank, the quantity bought are consistent.Outer packing meets the requirements and without breakage.Animal subject outer packing is through 75% wine
Smart spraying disinfection, after ultraviolet light irradiation 3 minutes by the incoming quarantine room of passing cabinet.
It examines: in the outer packing of chamber opening animal subject, animal subject gender and quantity and the animal subject verification of conformity of quarantining
Contained item is consistent.It is numbered and is marked in mouse tail root with marker pen, then animal subject is checked one by one, including checked
Gender, weight, head, trunk, tail portion, four limbs, fur, spirit and activity etc., all animal subjects are showed no obvious abnormalities.By
It is put into animal subject rearging cage after examination animal test, sub-cage rearing, every cage 3-5, and the hang tag in cage tool, is placed on inspection
Laundering period raising is carried out in epidemic disease room, all animal subjects are showed no exception in inspection.
(2) test sample is prepared:
Test sample preparation method: Normal group (control) directly takes commercially available 0.9% sodium chloride injection, is not necessarily to
It prepares；The human umbilical cord mesenchymal stem cells that MSC injection group uses, which are sent, to be used.
Test sample saves: umbilical cord mesenchymal stem cells are temporarily stored into 2-8 DEG C of ice chest, in 8 hours effectively if do not used immediately.
Test the processing of remaining test sample: test sample is cell product, places -20 DEG C of ice after remaining test sample high pressure inactivation
In cabinet, it is uniformly processed.
Administration: mouse single intravenous injection, tail vein are slowly injected.
(3) Index for examination
(I) mouse behavior observation: in raising, the situation of routine observation mouse weekly, and at least weighing is primary, and records
Weight.
Whether nodosity is formed (II) internal organs: after injecting MSC, when breeding cycle reaches 10 weeks and 6 months, passing through cervical vertebra
The method of dislocation puts to death mouse, and dissects.Observing main organs, whether nodosity is formed, and is taken pictures.
(III) organ weights and organ coefficient statistics: when breeding cycle reaches 10 weeks and 6 months, each group mouse is taken after dissect
The heart, liver, spleen, lung, kidney, thymus gland reject the fat of each internal organs attachment, weigh each organ weights, and calculate organ coefficient (internal organs system
Number=organ weights/weight × 1000).It calculates acropetal coefficient (mg/g) (acropetal coefficient=organ weight/weight).
The detection of (IV) Histopathology: in order to observe inside internal organs whether tuberculous formation, to the main organs of mouse
The case where being separated, prepare paraffin section, observation each organ of mouse is dyed by HE.
Experimental result:
(I) control group and MSC injection group upon administration, in 6 months feeding times, no phenomena of mortality, the state of mind and
Autonomic activities are normal, and secretion and excreta also no abnormality seen, changes of weight trend is identical, Normal group and MSC injection
The mouse weight variation of group is as shown in FIG. 1A and 1B.
(II) Normal group and MSC injection group were administered 10 weeks and after 6 months, MSC injection group and Normal group battalion
Feeding situation is all good, and each internal organs are showed no macroscopic significant changes, as a result as shown in Fig. 2A, Fig. 2 B, Fig. 2 C and Fig. 2 D, respectively
A internal organs form, color and quality are normal, and the tubercle being visible by naked eyes is formed.
(III) organ weights and coefficients statistics show Normal group and MSC injection group in administration injection sodium chloride respectively
Injection and 2 × 10610 weeks and after six months after a/MSC only, mice organs coefficient there are no significant difference, as a result such as figure
3A, Fig. 3 B, Fig. 3 C, Fig. 3 D, Fig. 3 E, Fig. 4 A, Fig. 4 B, Fig. 4 C, shown in Fig. 4 D and Fig. 4 E.
The detection of (IV) Histopathology: Normal group and MSC injection group are administered 10 weeks and main organs exist after 6 months
There is no marked difference in histology, does not observe the formation of tubercle, as a result as fig. 5 a and fig. 5b yet.
Experiment conclusion: human umbilical cord mesenchymal stem cells are 2 × 106It is a/only dosage under will not to BALB/c Nude mouse
Tumor.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of evaluation method of mescenchymal stem cell preparation oncogenicity, which is characterized in that the evaluation method includes: that offer connects
By the animal subject that the mescenchymal stem cell preparation is administered, the tumorigenesis of mescenchymal stem cell is then evaluated according to pre-set level
Property；
The dosage of the animal subject for receiving the mescenchymal stem cell preparation administration is 0.5 × 106~5 ×
106Cells/ is only；
The pre-set level includes following index:
(a1) changes of weight；
(a2) organ weights and organ coefficient；
(a3) whether nodosity is formed in the animal subject body.
2. evaluation method according to claim 1, which is characterized in that it is dry thin to receive the mesenchyma with the animal subject
The time of born of the same parents' preparation administration, the weight of the animal subject is obtained weekly.
3. evaluation method according to claim 1, which is characterized in that it is dry thin to receive the mesenchyma with the animal subject
After the time of born of the same parents' preparation administration starts 10 weeks, the organ weights and organ coefficient of the animal subject are obtained；
With, after starting 6 months with the time that the animal subject receives mescenchymal stem cell preparation administration, obtain it is described by
Try the organ weights and organ coefficient of animal；
Preferably, the internal organs include one or more of the heart, liver, spleen, lung, kidney and thymus gland.
4. evaluation method according to claim 1, which is characterized in that it is dry thin to receive the mesenchyma with the animal subject
After the time of born of the same parents' preparation administration starts 10 weeks, check in the animal subject body whether tubercle occur；
With, after starting 6 months with the time that the animal subject receives mescenchymal stem cell preparation administration, check it is described by
Whether there is tubercle in examination animal body；
Preferably, described to check that whether occurring tubercle in the animal subject body includes following (b1) and/or (b2):
(b1) it is dissected after putting to death animal subject, whether nodosity is formed observation internal organs；
(b2) the progress Histopathology detection of animal subject internal organs is obtained after putting to death animal subject.
5. evaluation method according to claim 1, which is characterized in that the evaluation method further includes blank control group；Institute
Stating blank control group includes the animal subject for receiving saline administration；
Preferably, the animal subject that the blank control group and the receiving mescenchymal stem cell preparation are administered is independently
Include 3~5 animal subjects.
6. evaluation method according to any one of claims 1-5, which is characterized in that the animal subject include mouse,
One or more of rat, cavy, rabbit, dog, miniature pig, monkey, ferret, marmot and naked mole；Preferably mouse.
7. evaluation method according to claim 6, which is characterized in that the animal subject is BALB/c Nude mouse；Institute
Stating BALB/c Nude mouse is 6-8 week old, and weight is the female mice of 18~20g.
8. evaluation method according to claim 7, which is characterized in that the dosage of the BALB/c Nude mouse is 1
×106~2.5 × 106Cells/ is only；Preferably 2 × 106Cells/ is only.
9. evaluation method according to claim 1, which is characterized in that the administration mode of the administration include subcutaneous administration,
Intramuscular administration, intravenous administration, intravaginal administration, intrauterine administration, pulmonary administration or rectally；
Preferably, the administration mode includes intravenous administration；More preferably mouse tail vein input administration.
10. the evaluation method of mescenchymal stem cell preparation oncogenicity of any of claims 1-9 is between preparation includes
Application in the drug of mesenchymal stem cells；
Preferably, the drug includes human umbilical cord mesenchymal stem cells.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811504926.9A CN109529054A (en) | 2018-12-10 | 2018-12-10 | The evaluation method and application of mescenchymal stem cell preparation oncogenicity |
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