CN115944651A - Application of human umbilical cord mesenchymal stem cells in preparation of medicine for treating secondary damage caused by cerebral artery occlusion infarction - Google Patents
Application of human umbilical cord mesenchymal stem cells in preparation of medicine for treating secondary damage caused by cerebral artery occlusion infarction Download PDFInfo
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Abstract
The invention discloses application of human umbilical cord mesenchymal stem cells in preparing a medicament for treating secondary damage caused by cerebral artery occlusion and infarction. The invention achieves the effect of repairing secondary damage in a far-separated brain area caused by middle cerebral artery occlusion infarction by preparing the medicament containing the human umbilical cord mesenchymal stem cells. The invention discovers that the human umbilical cord mesenchymal stem cells can relieve secondary damage generated at a far-septate substantia nigra part of an infarction focus of a cerebral cortex and repair dopaminergic neurons. The medicine of the invention not only improves the deletion of movement and sensory behavior after the middle cerebral artery occlusion infarction, but also improves the damage loss of dopaminergic neuron in the far brain area of the infarction focus after the infarction of one side cortex, and lays an important scientific basis for clinical treatment of secondary damage caused by the middle cerebral artery occlusion infarction by the human umbilical cord mesenchymal stem cells.
Description
Technical Field
The invention belongs to the technical field of stem cell therapy, and particularly relates to application of human umbilical cord mesenchymal stem cells in preparation of a medicine for treating secondary damage caused by middle cerebral artery occlusion infarction.
Background
Cerebrovascular disease is a common clinical disease with high disabling fatality rate, is the third leading cause of death in the population of China, and is also the first cause of death in all vascular diseases. With the aging of population, the incidence of ischemic cerebrovascular disease is on the trend of increasing year by year, and the health and life of human beings are seriously harmed. However, parkinsonism accompanying cerebral infarction has become one of the important causes of the difficulty in prognosis in elderly patients. Therefore, the research on the pathogenesis of the secondary parkinsonism after cerebral infarction has become a problem which needs to be solved urgently in the medical field.
In addition to ischemic necrosis of the ipsilateral cerebral cortex and striatum after Middle Cerebral Artery Occlusion (MCAO) in adult rats, secondary injury is also successively generated in the substantia nigra part far away from the striatum of the cortex on the same side of an infarction focus, and is characterized in that the substantia nigra reticulata part on the same side of the infarction is reduced in volume and neurons are greatly degenerated and lost. Since degenerative loss of nigral dopaminergic neurons is a major pathological feature of parkinsonism, degenerative damage to the distal nigral dopaminergic neurons following one-sided MCAO is an important cause of the induction of secondary parkinsonism.
Human umbilical cord mesenchymal stem cells are a cell population with the ability to self-renew, highly proliferate and multi-lineage differentiate. In the related art, the mode of treating cerebral apoplexy by using human umbilical cord mesenchymal stem cells and additives is mainly used for repairing damaged nerves around an infarct focus by reducing the size of the infarct focus of a cortical infarct model, reducing inflammatory reaction and the like. However, the current mechanism of treating cerebral apoplexy by human umbilical cord mesenchymal stem cells is only limited to repairing damaged neurons around cortical infarct focus and relieving inflammatory reaction, and no description is provided for secondary damage occurring at the far septal substantia nigra part.
Therefore, the field still needs to research the action mechanism of the human umbilical cord mesenchymal stem cells in the treatment of secondary damage in the far-separated brain area caused by the middle cerebral artery occlusion infarction deeply, and further utilize the characteristics to better exert the treatment value of the damage caused by the middle cerebral artery occlusion infarction.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides the application of the human umbilical cord mesenchymal stem cells in preparing the medicine for treating secondary damage caused by artery occlusion infarction in the brain. In the invention, the inventor finds that secondary damage occurring at a far-separated brain region part of a middle cerebral artery occlusion infarction focus can be relieved and repaired through the human umbilical cord mesenchymal stem cells, thereby providing a new idea and a new treatment means for secondary damage caused by middle cerebral artery occlusion.
In a first aspect of the invention, there is provided use of human umbilical cord mesenchymal stem cells in the manufacture of a medicament for the treatment of secondary damage caused by an occlusive infarction of the middle cerebral artery.
In some embodiments of the invention, the secondary damage is secondary damage to distant brain regions.
In some embodiments of the present invention, the method for preparing the human umbilical cord mesenchymal stem cell comprises the following steps:
extracting the purchased human umbilical cord mesenchymal stem cells, culturing in a special culture medium for the human umbilical cord mesenchymal stem cells, changing liquid for multiple times to remove non-adherent cells, culturing, subculturing and amplifying, then using for transplantation, digesting with trypsin with the final concentration of 0.25% before transplantation, blowing, beating and dispersedly culturing to obtain the human umbilical cord mesenchymal stem cells, diluting the trypsin with the special culture medium for the human umbilical cord mesenchymal stem cells until digestion is stopped, centrifuging, and then removing supernatant, thus obtaining the human umbilical cord mesenchymal stem cells.
In some embodiments of the invention, the human umbilical cord mesenchymal stem cells are purchased from Tianjin Angiosu cell Gene engineering, inc.
In some embodiments of the invention, the medium dedicated to human umbilical cord mesenchymal stem cells is mesenchymal stem cell medium purchased from Tianjin Angelen cell genetic engineering, inc.
In some embodiments of the present invention, the culture condition of the human umbilical cord mesenchymal stem cells is 35 to 37 ℃.
In some embodiments of the invention, the human umbilical cord mesenchymal stem cells are cultured in a medium specific for human umbilical cord mesenchymal stem cells, the composition of which comprises DMEM,10% fetal bovine serum and 5% double antibody (streptomycin cocktail).
In some embodiments of the present invention, the number of culture passages of the human umbilical cord mesenchymal stem cell is 8 to 11 passages.
In some embodiments of the present invention, the number of passages of culturing the human umbilical cord mesenchymal stem cells is 10 to 11 passages.
In some embodiments of the invention, the centrifugation is performed at 1000rpm,26 ℃ and 3min.
In some embodiments of the invention, the cell concentration of the human umbilical cord mesenchymal stem cells in the medicament is 1.0 × 10 6 ～3.0×10 6 Individual cells/mL.
In some embodiments of the invention, the medicament further comprises other pharmaceutically acceptable adjuvants.
In some embodiments of the invention, the pharmaceutically acceptable adjuvants include diluents, absorbents, wetting agents, binders, disintegrants, lubricants, colorants, coating materials, solvents, pH adjusters, antibacterial agents, isotonicity adjusters, chelating agents.
In some embodiments of the invention, the drug is an infusion agent.
In some embodiments of the invention, the mode of administration of the drug comprises subcutaneous injection, intradermal injection, intramuscular injection, intravenous drip, intrathecal injection.
In some embodiments of the invention, the drug is administered at a dose of 1.0X 10 6 ～3.0×10 6 One cell/rat.
In some embodiments of the present invention, the dosage of the drug can be reasonably obtained from dose conversion relationship between experimental animals, and other animals including, but not limited to, human, mouse, dog, pig, etc.
In some embodiments of the invention, the time node of administration of the drug is: the injection is injected for 1 time within 20 to 28 hours after the middle cerebral artery occlusion.
In some embodiments of the invention, the time node for administration of the drug is: the injection is injected 1 time within 24 hours after the middle cerebral artery occlusion.
In some embodiments of the invention, the drug further comprises other neuronal repair drugs.
In some embodiments of the invention, the other neuron repair drugs comprise edaravone, gangliosides, citicoline, cerebrolysin, oxiracetam.
The invention is proved by the experiments of the embodiment: the human umbilical cord mesenchymal stem cells can improve the loss of movement and sensory behaviors after the cerebral middle artery occlusion infarction and simultaneously improve the damage loss of dopaminergic neurons in a far brain area of an infarction focus after the cerebral middle artery occlusion infarction.
The invention has the beneficial effects that:
1. the invention discovers that the human umbilical cord mesenchymal stem cells can repair secondary damage in a far-separated brain area caused by middle cerebral artery occlusion infarction.
2. The invention not only discovers that the human umbilical cord mesenchymal stem cells can improve the loss of movement and sensory behavior after the middle cerebral artery occlusion infarction, but also improves the loss of neurons in the far brain separating area of the infarct focus after the infarction of the cortex on one side.
3. The invention establishes a method for repairing secondary damage in a far-separated brain area caused by cerebral cortex infarction and cerebral artery occlusion infarction by using human umbilical cord mesenchymal stem cells, and lays an important scientific basis for clinical treatment of secondary damage caused by cerebral artery occlusion infarction by using human umbilical cord mesenchymal stem cells.
Drawings
The invention is further described with reference to the following figures and examples, in which:
FIG. 1 is a flow chart of an animal model experiment according to an embodiment of the present invention;
FIG. 2 is a diagram of a pattern of secondary damage in a far-septal area caused by an occlusive infarction of a middle cerebral artery, wherein A is a diagram of an occlusive coronary section of the middle cerebral artery, B is a diagram of a coronary section of the secondary damage in the far-septal area, and in the diagrams, the area (1) is an ischemic area after the occlusive injury of the middle cerebral artery and the area (2) is a part where the secondary damage in the far-septal area occurs;
FIG. 3 shows the results of a rat gummed paper pasting experiment according to an embodiment of the present invention;
FIG. 4 is a graph showing the evaluation of neurological deficit in rats according to an example of the present invention;
FIG. 5 is a photograph of NeuN and TH immunohistochemical staining of secondary lesions in the far brain area caused by infarction of middle cerebral artery occlusion in an example of the present invention, wherein the a-d plots are: TH immunohistochemical staining pattern; the e-h diagram is as follows: neuN immunohistochemical staining profile; a. e is blank control group; b. f is a model control group; c. g is low dose group; d. h is high dose group;
fig. 6 is a graph showing the statistics of TH positive cell counts of secondary lesions in the distal cerebral region caused by infarction, wherein left side closure injury is the number of cells measured after left side cerebral artery occlusion in the middle brain, and right side non-injury is the number of cells measured without right side cerebral occlusion in the middle brain, which indicates that p is less than 0.05 when compared with the blank control group, # is compared with the model control group black;
fig. 7 is a statistical graph of the NeuN positive cell count of a substantia nigra brain region of secondary damage to a distal substantia nigra brain region caused by an occlusive infarction of an artery in the middle brain in an example of the present invention, wherein the left side is the cell count of the distal substantia nigra brain region on the same side after left-side middle artery occlusion, and the right side is intact and is the cell count of the substantia nigra brain region on the right side of the brain, # is compared to a blank control group and # is less than 0.05 compared to a model control group.
Detailed Description
The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.
The test materials and reagents used in the examples of the present invention are all consumable materials and reagents that are conventionally available from commercial sources unless otherwise specified.
Preparation of human umbilical cord mesenchymal stem cells
In the embodiment of the invention, the human umbilical cord mesenchymal stem cells are purchased from Tianjin Oncai cell gene engineering Co.
In the embodiment of the invention, the inventor carries out activation culture on the purchased human umbilical cord mesenchymal stem cells, and then the obtained cells are treated and then applied to transplantation, and the specific steps are as follows:
culturing human umbilical cord mesenchymal stem cells in a special culture medium (mesenchymal stem cell culture medium purchased from Tianjin Oncai cell genetic engineering Co., ltd.) for the human umbilical cord mesenchymal stem cells, changing the medium for multiple times to remove the cells which are not attached to the wall, culturing for passage expansion for 10-11 times at 37 ℃, digesting and blowing dispersed cells by trypsin with the final concentration of 0.25% before cell transplantation after the cells grow to 90% of the visual field, diluting the trypsin by the special culture medium for the human umbilical cord mesenchymal stem cells until digestion is stopped, centrifuging the dispersed cells for 3min at 1000rpm and 26 ℃, and discarding the supernatant to obtain the human umbilical cord mesenchymal stem cells. Repair effect of human umbilical cord mesenchymal stem cells on secondary damage of far-separated brain area caused by middle cerebral artery occlusion infarction
In the embodiment of the invention, the inventor adopts an animal experiment mode to verify the repair effect of the human umbilical cord mesenchymal stem cells on secondary damage of a far-separated brain area caused by middle cerebral artery occlusion infarction.
In the embodiment of the invention, the tested animals are 24 male Sprague-Dawley rats (SD rats) with the weight of 240-280 g purchased from the centers of animals in Guangdong province, all the SD rats for experiments are arranged under standard laboratory conditions, the 24 male SD rats are randomly divided into 4 groups including a blank control group (a sham operation group), a model control group, a low dose group and a high dose group after being adapted to the environment by free drinking water and eating for 7 days, the experimental environment is controlled to be 21-23 ℃ at room temperature and the relative humidity is controlled to be 40-50%, the light and shade circulation is carried out according to the period of 12-hour light and 12-hour dark, and the indoor light intensity is controlled to be 10-20 lux.
The specific experimental steps are as follows:
experimental SD rats were taken to be familiar with the normal diet at room temperature and 12h light/dark cycle for 7 days of acclimation. Then 24 SD rats in 4 groups of model control group, low dose group and high dose group are subjected to left Cerebral cortex Middle Artery Occlusion infarction operation by electrocoagulation to construct a left Middle Cerebral Artery embolism (MCAO) model, while rats in a blank control group (pseudo operation group) are only subjected to anesthesia craniotomy and are not subjected to Occlusion infarction operation. The behavioural tests were performed at 1,7, 14, 21, 28 days post-surgery, during which four groups were supplied with the same normal feed and water according to the normal feeding standards of SD rats, and after completion of the behavioural tests at day 28, the rats were euthanized and 6 rats per group were randomly selected and brain slices were taken for immunohistochemical testing. The experimental procedure is shown in FIG. 1.
The operation of cerebral cortex middle artery occlusion infarction of the experiment comprises the following specific steps: rats were fasted for 12 hours before surgery, anesthetized with 10% chloral hydrate, and the midpoint of the eye-ear line was cut perpendicular to it, and exposed to the skull layer by layer. Drilling a bone window with the diameter of 5mm at the front lower part of the joint of the zygomatic bone of the left brain and the temporal bone scale part, carefully tearing off the dura mater by using an iris hook, closing the branch part of the cortex of the MCA by using bipolar electric coagulation forceps at the position between the olfactory tract and the Superior Cerebral Vein (SCV) and above the intersection of the Middle Cerebral Artery (MCA) and the SCV, suturing the incision, disinfecting the skin, and paying attention to infection resistance after the operation until the rat is awake. Post-operatively, the region of whole blood with occluded middle cerebral artery appears in section (1) of fig. 2.
The specific treatment conditions in each group were:
(1) Blank control group (sham group): anesthesia craniotomy only, without using bipolar coagulation forceps to occlude the branch site of the MCA cortex, and at 24 hours after surgery, 1mL of 0.01M Phosphate Buffered Saline (PBS) containing disodium hydrogenphosphate, sodium dihydrogenphosphate and sodium chloride was slowly injected into the tail vein of the rat;
(2) Model control group: performing middle cerebral artery occlusion infarction operation, and slowly injecting 1mL of 0.01M PBS containing disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride into tail vein of rat at 24h after the operation;
(3) Low dose group: performing artery occlusion infarction operation in cerebral cortex, and adjusting human umbilical cord mesenchymal stem cell concentration to 1.0 × 10 with 0.01M PBS containing disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride at 24 hr after operation 6 1mL of human umbilical cord mesenchymal stem cells with well-adjusted cell concentration are slowly injected into tail vein of a rat per mL;
(4) High dose group: performing artery occlusion infarction operation in cerebral cortex, and adjusting human umbilical cord mesenchymal stem cell concentration to 3.0 × 10 by using 0.01M PBS containing disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride at 24 hr after operation 6 1mL of human umbilical cord mesenchymal stem cells with well-adjusted cell concentration is slowly injected into tail vein of rat per mL.
And (3) performing behavioral tests on rats of a model control group, a low-dose group and a high-dose group at 1,7, 14, 21 and 28 days after the arterial occlusive infarction in the cerebral cortex, and verifying the repairing effect of the human umbilical cord mesenchymal stem cells, wherein the blank control group is a pseudo-operation group, does not make a cerebral middle artery occlusive model, does not have the condition of behavioral deletion, and therefore is not included in the score.
The specific ethological test in the embodiment is a gummed paper pasting experiment, and the nerve function defect scoring is performed on an experimental rat, and the specific steps are as follows:
(1) Gummed paper sticking experiment:
placing all rats for testing in separate cages before testing, wherein a 200 mm piece of adhesive tape is adhered to the front paw on the opposite side of the post-operative infarction during the adhesion experiment; the test measures the time required for the rat to remove the sticker within 2 minutes, during which the rat pulls off the sticker from the forepaw in order to eliminate the irritation of the forelimb. Rats were kept in separate cages for testing during the test; the better the recovery of sensory function in the rat, the shorter the time to tear off the gummed paper.
The results are shown in table 1 and fig. 3.
TABLE 1SD rat time table for removal of stickies
(2) Neurological Sensory Scopes (NSS):
the Neurological deficit score (NSS) used in this experiment was improved and then tested for scoring, and the specific procedures were as follows: the tail was lifted and the degree of contralateral bending of the rat was observed. Rats were placed on the cage lid and the tail pulled back to test the gripping ability of the rat forepaws. Compared with the unmodified walking beam balance test scoring, the improved ethological scoring is simple and easy to operate, and the scoring standard is as follows:
10 minutes: no manifestation of neurological deficit;
8 min: the contralateral forepaw can not be fully extended;
6 min: buckling the contralateral forepaw;
and 4, dividing: turning to the outside;
and 2, dividing: when walking, the walking stick is inclined towards the opposite side;
0 minute: inability to walk and loss of consciousness.
Animals were scored for neurological function after waking, with scores of 4-9 being included in the study and scores of 0 and 10 being excluded. The exercise test criteria were: lifting the tail part, and observing the bending degree of the rat to the affected side; the index is 0 point for no abnormality, 7-8 points for medium abnormality and 9 points for serious abnormality.
The results are shown in table 2 and fig. 4.
TABLE 2 improved neurological functionality scoring table
| Group of | |
|
|
|
Day 28 |
| Model control group | 8.6 | 7.4 | 7 | 6.8 | 5.9 |
| Low dose group | 8.6 | 6.2 | 5.2 | 4.3 | 3.4 |
| High dose group | 8.5 | 7.1 | 6.1 | 5 | 4.3 |
As shown in table 2 and fig. 4, the lack of sensory and motor functions was found to be improved in both the low dose and high dose groups compared to the model control group without stem cell injection, and the difference was statistically significant in both the gummed paper adhesion test and the neurological scoring. The low dose group was not significantly different from the high dose group, considering that the dose of human mesenchymal stem cells may not have significant impact on the behavioral improvement.
After the behavioral test is finished, taking pathological tissue materials and immunohistochemical staining are carried out, and the method comprises the following specific steps:
(1) Taking pathological tissues: at 28 days post-surgery time points in the placebo (sham), model, low and high dose groups, 6 subjects were sacrificed at random for brain slice preparation. After deep anesthesia, a proper amount of normal saline is perfused through an ascending aorta, then a proper amount of 4% paraformaldehyde and 0.1M PBS (pH value of 7.4) are perfused rapidly, the cranium is rapidly opened to take out the brain, 4% paraformaldehyde and 0.1M PBS (pH value of 7.4) are continuously fixed for 24 hours, and after dehydration is carried out in three sucrose solutions with gradient concentrations of 10%, 20% and 30% in sequence until the brain is settled, frozen sections are carried out at-10 ℃ and the thickness of the slices is 30 micrometers. Brain pieces were stored in 0.1M PBS for immunohistochemical staining detection.
(2) Immunohistochemical staining: the brain slices prepared in the blank control group, the model control group, the low-dose group and the high-dose group are respectively subjected to immunohistochemical research experiments. Brain pieces from bregma4.52 to 6.3mm containing substantia nigra parts were taken, treated with 0.01M PBS, 3% hydrogen peroxide solution for 30 minutes, then treated with 5% blocking for 1 hour with normal rat serum at room temperature, and then treated separately with different primary antibodies, both rabbit polyclonal anti-Tyrosine Hydroxylase (TH) (dilution 1: 5000) and mouse monoclonal antibody anti-neuron specific nuclear binding protein (NeuN) (dilution 1: 2000), which were purchased from millipore sigma company. Incubate overnight at 4 ℃ for 12-16h. After completion of incubation, the cells were washed three times with 0.01M PBS, incubated with immunoglobulin G (IgG) as a secondary antibody at room temperature for 2 hours, washed with 0.01M PBS, sliced, and incubated with ABC peroxidase at room temperature for 30 minutes. The brain slices prepared in the experimental process are dyed by a diaminobenzidine method, and the peroxidase DAB color reaction of the brain slices containing 0.05 percent of diaminobenzidine and 0.01 percent of hydrogen peroxide is observed. 3 non-overlapping parts of the ipsilateral and contralateral Substantia nigra pars compacta (SNc) of each brain section were selected and photographed under an optical microscope of 200 times to prepare an immunohistochemical staining image, which is shown in FIG. 5 and is specifically taken from the (2) region in the B image of FIG. 2, i.e., the occurrence part of secondary damage in the distant brain region; the numbers of NeuN, TH positive immune cells were calculated using ImageJ's cell counting software (NIH, bethesda, MD, usa) as shown in fig. 6, 7. The data were determined in this experiment under double-blind conditions.
As can be seen from the immunohistochemical staining chart of FIG. 5, the number of cells in the low-dose and high-dose groups was increased compared to the model control group, which demonstrates that human umbilical cord mesenchymal stem cells have neural repair effects on dopaminergic neurons and all neural pronuclei of substantia nigra to different degrees.
From the statistical results of the number of positive cells of the dopaminergic neurons (TH) in fig. 6 and the neuronal nuclear marker (NeuN) in fig. 7, it can be seen that the number of markers in the left substantia nigra region and the right substantia nigra region of the brain after the middle cerebral artery occlusion infarct surgery is reduced relative to the blank control group, wherein the number of positive cells of the markers is increased in the low dose and high dose groups compared with the model control group, which indicates that both doses have different degrees of nerve repair, and the difference has statistical significance; the number of positive cells of the two dose groups is further analyzed, the low dose group has no significant difference compared with the high dose group, and the dosage of the human umbilical cord mesenchymal stem cells is considered to have no significant influence on nerve cell repair.
All control data in this example are expressed as mean standard deviation; and Statistical analysis was performed using SPSS 13.0 (Statistical Product Service Solutions, SPSS, inc., chicago, il, usa); significance was determined by two-tailed t-test or one-way test; analysis of variance, then LSD (Least-SignificantDifference) or Tamhane's T2 post hoc test analysis; p <0.05 was statistically significant.
The results show that the human umbilical cord mesenchymal stem cells can be effectively applied to the application of repairing secondary damage in a far-separated brain area caused by arterial occlusive infarction in the brain, and the repairing effect is excellent. Whether the neuron repair condition analysis of the behavioral or pathological section is carried out, the fact that the human umbilical cord mesenchymal stem cells repair the behavioral and nerve injuries in different degrees can be observed, the experimental result of the embodiment suggests that the human umbilical cord mesenchymal stem cells have a treatment effect on cerebral apoplexy and possibly brought secondary Parkinson, and theoretical basis is provided for clinically treating the far septal cerebral region secondary damage caused by the cerebral middle artery occlusion infarction by the human umbilical cord mesenchymal stem cells.
The embodiments of the present invention have been described in detail with reference to the accompanying drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present invention. Furthermore, the embodiments of the present invention and the features of the embodiments may be combined with each other without conflict.
Claims (10)
1. The application of the human umbilical cord mesenchymal stem cells in preparing the medicine for treating secondary damage caused by artery occlusion infarction in the brain.
2. The use of claim 1, wherein the secondary damage is secondary damage to a distant brain region.
3. The use according to claim 1, wherein the number of culture passages of the human umbilical cord mesenchymal stem cells is 8-11 generations.
4. The use of claim 1, wherein the cell concentration of human umbilical cord mesenchymal stem cells in the medicament is 1.0 x 10 6 ～3.0×10 6 Individual cells/mL.
5. The use of claim 1, wherein the medicament further comprises other pharmaceutically acceptable adjuvants.
6. The use of claim 5, wherein the pharmaceutically acceptable adjuvants comprise diluents, absorbents, wetting agents, binders, disintegrants, lubricants, colorants, coating materials, solvents, pH adjusters, antimicrobials, isotonicity adjusters, chelating agents.
7. The use according to claim 1, wherein the medicament is administered by subcutaneous injection, intradermal injection, intramuscular injection, intravenous drip, intrathecal injection.
8. The use of claim 1, wherein the medicament is administered at a dose of 1.0 x 10 6 ～3.0×10 6 One cell/rat.
9. The use of claim 1, wherein the time node for administration of the drug is: the injection is injected for 1 time within 20 to 28 hours after the middle cerebral artery occlusion.
10. The use of claim 1, wherein the medicament further comprises an additional neuronal repair agent.
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| CN116999465A (en) * | 2023-09-19 | 2023-11-07 | 河北中琨生物工程有限责任公司 | Application of umbilical cord mesenchymal stem cell-derived exosome in preparation of medicines for preventing and/or treating ischemic cerebral apoplexy |
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| CN116999465A (en) * | 2023-09-19 | 2023-11-07 | 河北中琨生物工程有限责任公司 | Application of umbilical cord mesenchymal stem cell-derived exosome in preparation of medicines for preventing and/or treating ischemic cerebral apoplexy |
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