CN108542917A - A kind for the treatment of rheumatic ostealgia disease treatment injection and preparation method thereof with person joint's liquid mescenchymal stem cell excretion body extract - Google Patents
A kind for the treatment of rheumatic ostealgia disease treatment injection and preparation method thereof with person joint's liquid mescenchymal stem cell excretion body extract Download PDFInfo
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- 239000007788 liquid Substances 0.000 title claims abstract description 179
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 167
- 230000029142 excretion Effects 0.000 title claims abstract description 115
- 239000000284 extract Substances 0.000 title claims abstract description 85
- 238000002347 injection Methods 0.000 title claims abstract description 71
- 239000007924 injection Substances 0.000 title claims abstract description 71
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 48
- 208000025747 Rheumatic disease Diseases 0.000 title claims abstract description 42
- 230000000552 rheumatic effect Effects 0.000 title claims abstract description 42
- 206010006002 Bone pain Diseases 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 239000000243 solution Substances 0.000 claims abstract description 38
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 9
- 239000008103 glucose Substances 0.000 claims abstract description 9
- 210000004027 cell Anatomy 0.000 claims description 26
- 239000012141 concentrate Substances 0.000 claims description 22
- 238000005119 centrifugation Methods 0.000 claims description 18
- 239000001963 growth medium Substances 0.000 claims description 18
- 210000003716 mesoderm Anatomy 0.000 claims description 14
- 239000007853 buffer solution Substances 0.000 claims description 12
- 201000010099 disease Diseases 0.000 claims description 12
- 230000028327 secretion Effects 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 8
- 238000000746 purification Methods 0.000 claims description 8
- 238000003786 synthesis reaction Methods 0.000 claims description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- 239000013049 sediment Substances 0.000 claims description 6
- 230000001256 tonic effect Effects 0.000 claims description 6
- 210000000988 bone and bone Anatomy 0.000 claims description 5
- 208000027753 pain disease Diseases 0.000 claims description 5
- 230000000694 effects Effects 0.000 abstract description 3
- 239000012530 fluid Substances 0.000 description 5
- 238000000605 extraction Methods 0.000 description 4
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
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Abstract
Include following component the invention discloses a kind for the treatment of rheumatic ostealgia disease treatment injection with person joint's liquid mescenchymal stem cell excretion body extract：Person joint's liquid mescenchymal stem cell excretion body extract solution of 5% 20% concentration containing 500 μ l, 800 μ l in per 1ml injections, remaining for the glucose and 8.5% 9% concentration of 5% concentration physiological saline, the invention also discloses a kind of preparation method of the treating rheumatic ostealgia disease treatment injection with person joint's liquid mescenchymal stem cell excretion body extract, which includes the following steps：S1：Obtain person joint's liquid mescenchymal stem cell, S2：Cultivate person joint's liquid mescenchymal stem cell, S3：The rush of person joint's liquid mescenchymal stem cell excretion body point, S4：Person joint's liquid mescenchymal stem cell excretion body extracts, S5：Injection is synthesized, treatment of the treating rheumatic ostealgia disease treatment injection prepared by the present invention with respect to treating rheumatic ostealgia disease has the effect of highly significant and can improve the immunocompetence of human body, therefore has good market prospects, is suitble to promote.
Description
Technical field
The present invention relates to treating rheumatic ostealgia disease treatment injection preparing technical fields, specially a kind of between person joint's liquid
Treating rheumatic ostealgia disease treatment injection of mesenchymal stem cells excretion body extract and preparation method thereof and preparation method thereof.
Background technology
Mescenchymal stem cell (MSC, mesenchymal stem cells) is the important member of stem cell line, is derived from
The mesoderm of mesoderm growing early stage belongs to multipotential stem cell, and MSC initially has found in marrow, because it is with multi-lineage potential, hematopoiesis
Support and promote stem cell implantation, immunoregulation and the concern that people are increasingly subject to the features such as self-replacation；
And excretion body refers to the small film bubble (30-150nm) for containing complicated RNA and protein, now, refers in particular to diameter and exists
The plate-like vesica of 40-100nm can derive from a variety of different cells, and the excretion body that different cells generate includes difference
The excretion body of protein, fat and mRNA of type etc., each cell all have the function of itself unique immune, reparation etc.；
And the treatment of the treating rheumatic ostealgia disease of the ingredient counterpart contained by person joint's liquid mescenchymal stem cell excretion body has
Notable curative effect, for this purpose, the present invention proposes a kind for the treatment of rheumatic ostealgia with person joint's liquid mescenchymal stem cell excretion body extract
Disease treatment injection and preparation method thereof.
Invention content
The purpose of the present invention is to provide a kind of rheumatic bones with person joint's liquid mescenchymal stem cell excretion body extract
Pain disease treatment injection and preparation method thereof and preparation method thereof, to solve the problems mentioned in the above background technology.
To achieve the above object, the present invention provides the following technical solutions：One kind is with outside person joint's liquid mescenchymal stem cell
The treating rheumatic ostealgia disease treatment injection of body extract is secreted, it is described a kind of with the extraction of person joint's liquid mescenchymal stem cell excretion body
The treating rheumatic ostealgia disease treatment injection of object includes following component：5%-20% containing 500 μ l-800 μ l in per 1ml injections
Person joint's liquid mescenchymal stem cell excretion body extract solution of concentration, remaining is the life of 5% glucose and 8.5%-9% concentration
Manage brine.
Preferably, person joint's liquid mescenchymal stem cell excretion body extract solution is the concentration via centrifugation purification
Liquid.
Preferably, person joint's liquid mesenchyma used by person joint's liquid mescenchymal stem cell excretion body extract solution
Stem cell is the third generation to band six generations stem cell.
Preferably, the storage temperature of person joint's liquid mescenchymal stem cell excretion body extract solution is subzero 20
Degree.
Preferably, a kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract
Injection includes following component：Person joint's liquid mescenchymal stem cell excretion body containing 500 μ l, 5% concentration in per 1ml injections
Extract solution, remaining is the glucose of 5% concentration.
Preferably, a kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract
Injection includes following component：Person joint's liquid mescenchymal stem cell excretion containing 600 μ l, 10% concentration in per 1ml injections
Body extract solution, remaining is the physiological saline of 8.5% concentration.
Preferably, a kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract
Injection includes following component：Person joint's liquid mescenchymal stem cell excretion containing 700 μ l, 15% concentration in per 1ml injections
Body extract solution, remaining is the physiological saline of 9% concentration.
Preferably, a kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract
Injection includes following component：Person joint's liquid mescenchymal stem cell excretion containing 800 μ l, 20% concentration in per 1ml injections
Body extract solution, remaining is the physiological saline of 9% concentration.
A kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract as described above
The preparation method of injection, the preparation method include the following steps：
S1：Person joint's liquid mescenchymal stem cell is obtained, person joint's liquid of acquisition is launched in sterile centrifugation tube, and is added
PBS buffer solution is uniformly mixed, and is then centrifuged with cell Separating tube and is obtained cell separating liquid, supernatant liquor is removed, and will
Lower sediment thing is added in PBS buffer solution and washs, and person joint's liquid mescenchymal stem cell is obtained after centrifuging again；
S2：Person joint's liquid mescenchymal stem cell is cultivated, person joint's liquid mescenchymal stem cell of acquisition is put into culture dish
Culture, person joint's liquid mescenchymal stem cell of hair tonic is the first joint fluid mescenchymal stem cell in culture dish after culture,
By first generation person joint's liquid mescenchymal stem cell of acquisition obtained after five secondary cultures the second generation, the third generation, forth generation,
5th generation, the 6th generation person joint's liquid mescenchymal stem cell；
S3：The rush of person joint's liquid mescenchymal stem cell excretion body point selects the third generation to the 6th generation person joint's liquid mesenchyma
Stem cell is placed in the incubator that oxygen content is 25%, is cultivated using high speed culture medium, when cell bout degree is 75%,
It uses instead and the synthesis of excretion body and the culture medium of secretion is promoted to carry out culture 3-7 days；
S4：Person joint's liquid mescenchymal stem cell excretion body extracts, and will promote the culture medium of excretion body synthesis and secretion in S3
After carrying out culture 3-7 days, standing collects supernatant liquor i.e. person joint's liquid mescenchymal stem cell excretion body, then by the clear liquid of collection
Concentrate is obtained after centrifugation purifies, concentrate, which is statically placed in subzero 20 degree, to be sealed up for safekeeping；
S5：Injection is synthesized, the concentration of person joint's liquid mescenchymal stem cell excretion body extract of 5%-20% concentration is taken
Liquid obtains injection after being thoroughly mixed respectively with the glucose of the physiological saline of 8.5%-9% concentration and 5%.
Compared with prior art, the beneficial effects of the invention are as follows：Person joint's liquid mescenchymal stem cell is carried in the present invention
A large amount of person joint is cultivated using cell culture apparatus when the preparation of the treating rheumatic ostealgia disease treatment injection of excretion body extract
Liquid mescenchymal stem cell, and purification, this kind of extraction side are repeatedly concentrated again after generating excretion body by person joint's liquid mescenchymal stem cell
It is owned by France in biological extraction process, make injection that there is good bioactivity, and the treating rheumatic ostealgia disease prepared by the present invention
Treating treatment of the injection with respect to treating rheumatic ostealgia disease has the effect of highly significant, and the human body after injection is treated improves
Autoimmunity ability and disease resistance, therefore there are good market prospects, it is suitble to promote.
Specific implementation mode
The technical scheme in the embodiments of the invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is common
The every other embodiment that technical staff is obtained without making creative work belongs to the model that the present invention protects
It encloses.
Embodiment one
The present invention provides a kind of technical solution：A kind of rheumatism with person joint's liquid mescenchymal stem cell excretion body extract
Ostalgia disease treatment injection includes following component：Person joint's liquid mesenchyma containing 500 μ l, 5% concentration in per 1ml injections
Stem cell excretion body extract solution, remaining is the glucose of 5% concentration.
Further, person joint's liquid mescenchymal stem cell excretion body extract solution is the concentrate via centrifugation purification.
Further, person joint's liquid mesenchyma used by person joint's liquid mescenchymal stem cell excretion body extract solution is dry
Cell is the third generation to band six generations stem cell.
Further, the storage temperature of person joint's liquid mescenchymal stem cell excretion body extract solution is subzero 20 degree.
A kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract as described above
The preparation method of injection, the preparation method include the following steps：
S1：Person joint's liquid mescenchymal stem cell is obtained, person joint's liquid of acquisition is launched in sterile centrifugation tube, and is added
PBS buffer solution is uniformly mixed, and is then centrifuged with cell Separating tube and is obtained cell separating liquid, supernatant liquor is removed, and will
Lower sediment thing is added in PBS buffer solution and washs, and person joint's liquid mescenchymal stem cell is obtained after centrifuging again；
S2：Person joint's liquid mescenchymal stem cell is cultivated, person joint's liquid mescenchymal stem cell of acquisition is put into culture dish
Culture, person joint's liquid mescenchymal stem cell of hair tonic is the first joint fluid mescenchymal stem cell in culture dish after culture,
By first generation person joint's liquid mescenchymal stem cell of acquisition obtained after five secondary cultures the second generation, the third generation, forth generation,
5th generation, the 6th generation person joint's liquid mescenchymal stem cell；
S3：The rush of person joint's liquid mescenchymal stem cell excretion body point selects third generation person joint's liquid mescenchymal stem cell to set
It in the incubator that oxygen content is 25%, is cultivated using high speed culture medium, when cell bout degree is 75%, uses promotion instead
Excretion body is synthesized carries out culture 3 days with the culture medium of secretion；
S4：Person joint's liquid mescenchymal stem cell excretion body extracts, and will promote the culture medium of excretion body synthesis and secretion in S3
After carrying out culture 3 days, standing collects supernatant liquor i.e. person joint's liquid mescenchymal stem cell excretion body, then passes through the clear liquid of collection
Concentrate is obtained after crossing centrifugation purification, concentrate, which is statically placed in subzero 20 degree, to be sealed up for safekeeping；
S5：Synthesize injection, take the concentrate of person joint's liquid mescenchymal stem cell excretion body extract of 5% concentration with
The glucose of 5% concentration obtains injection after being thoroughly mixed.
Embodiment two
The present invention provides a kind of technical solution：A kind of rheumatism with person joint's liquid mescenchymal stem cell excretion body extract
Ostalgia disease treatment injection includes following component：It is filled between person joint's liquid containing 600 μ l, 10% concentration in per 1ml injections
Matter stem cell excretion body extract solution, remaining is the physiological saline of 8.5% concentration.
Further, person joint's liquid mescenchymal stem cell excretion body extract solution is the concentrate via centrifugation purification.
Further, person joint's liquid mesenchyma used by person joint's liquid mescenchymal stem cell excretion body extract solution is dry
Cell is the third generation to band six generations stem cell.
Further, the storage temperature of person joint's liquid mescenchymal stem cell excretion body extract solution is subzero 20 degree.
A kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract as described above
The preparation method of injection, the preparation method include the following steps：
S1：Person joint's liquid mescenchymal stem cell is obtained, person joint's liquid of acquisition is launched in sterile centrifugation tube, and is added
PBS buffer solution is uniformly mixed, and is then centrifuged with cell Separating tube and is obtained cell separating liquid, supernatant liquor is removed, and will
Lower sediment thing is added in PBS buffer solution and washs, and person joint's liquid mescenchymal stem cell is obtained after centrifuging again；
S2：Person joint's liquid mescenchymal stem cell is cultivated, person joint's liquid mescenchymal stem cell of acquisition is put into culture dish
Culture, person joint's liquid mescenchymal stem cell of hair tonic is the first joint fluid mescenchymal stem cell in culture dish after culture,
By first generation person joint's liquid mescenchymal stem cell of acquisition obtained after five secondary cultures the second generation, the third generation, forth generation,
5th generation, the 6th generation person joint's liquid mescenchymal stem cell；
S3：The rush of person joint's liquid mescenchymal stem cell excretion body point selects forth generation person joint's liquid mescenchymal stem cell to set
It in the incubator that oxygen content is 25%, is cultivated using high speed culture medium, when cell bout degree is 75%, uses promotion instead
Excretion body is synthesized carries out culture 4 days with the culture medium of secretion；
S4：Person joint's liquid mescenchymal stem cell excretion body extracts, and will promote the culture medium of excretion body synthesis and secretion in S3
After carrying out culture 4 days, standing collects supernatant liquor i.e. person joint's liquid mescenchymal stem cell excretion body, then passes through the clear liquid of collection
The concentrate for obtaining person joint's liquid mescenchymal stem cell excretion body extract after centrifugation purifies again is crossed, concentrate is statically placed in zero
Lower 20 degree are sealed up for safekeeping；
S5：Synthesize injection, take the concentrate of person joint's liquid mescenchymal stem cell excretion body extract of 10% concentration with
The physiological saline of 8.5% concentration obtains injection after being thoroughly mixed.
Embodiment three
The present invention provides a kind of technical solution：It is described a kind of with person joint's liquid mescenchymal stem cell excretion body extract
Treating rheumatic ostealgia disease treatment injection includes following component：Person joint's liquid containing 700 μ l, 15% concentration in per 1ml injections
Mescenchymal stem cell excretion body extract solution, remaining is the physiological saline of 9% concentration.
Further, person joint's liquid mescenchymal stem cell excretion body extract solution is the concentrate via centrifugation purification.
Further, person joint's liquid mesenchyma used by person joint's liquid mescenchymal stem cell excretion body extract solution is dry
Cell is the third generation to band six generations stem cell.
Further, the storage temperature of person joint's liquid mescenchymal stem cell excretion body extract solution is subzero 20 degree.
A kind for the treatment of rheumatic ostealgia disease treatment with person joint's liquid mescenchymal stem cell excretion body extract as described above
The preparation method of injection, the preparation method include the following steps：
S1：Person joint's liquid mescenchymal stem cell is obtained, person joint's liquid of acquisition is launched in sterile centrifugation tube, and is added
PBS buffer solution is uniformly mixed, and is then centrifuged with cell Separating tube and is obtained cell separating liquid, supernatant liquor is removed, and will
Lower sediment thing is added in PBS buffer solution and washs, and person joint's liquid mescenchymal stem cell is obtained after centrifuging again；
S2：Person joint's liquid mescenchymal stem cell is cultivated, person joint's liquid mescenchymal stem cell of acquisition is put into culture dish
Culture, person joint's liquid mescenchymal stem cell of hair tonic is the first joint fluid mescenchymal stem cell in culture dish after culture,
By first generation person joint's liquid mescenchymal stem cell of acquisition obtained after five secondary cultures the second generation, the third generation, forth generation,
5th generation, the 6th generation person joint's liquid mescenchymal stem cell；
S3：The rush of person joint's liquid mescenchymal stem cell excretion body point selects the 5th generation person joint's liquid mescenchymal stem cell to set
It in the incubator that oxygen content is 25%, is cultivated using high speed culture medium, when cell bout degree is 75%, uses promotion instead
Excretion body is synthesized carries out culture 6 days with the culture medium of secretion；
S4：Person joint's liquid mescenchymal stem cell excretion body extracts, and will promote the culture medium of excretion body synthesis and secretion in S3
After carrying out culture 6 days, standing collects supernatant liquor i.e. person joint's liquid mescenchymal stem cell excretion body, then passes through the clear liquid of collection
The concentrate for obtaining person joint's liquid mescenchymal stem cell excretion body extract after centrifugation purifies again is crossed, concentrate is statically placed in zero
Lower 20 degree are sealed up for safekeeping；
S5：Injection is synthesized, the concentrate of person joint's liquid mescenchymal stem cell excretion body extract of 15% concentration is taken
Injection is obtained after being thoroughly mixed with the physiological saline of 9% concentration.
Example IV
The present invention provides a kind of technical solution：A kind of rheumatism with person joint's liquid mescenchymal stem cell excretion body extract
Ostalgia disease treatment injection includes following component：It is filled between person joint's liquid containing 800 μ l, 20% concentration in per 1ml injections
Matter stem cell excretion body extract solution, remaining is the physiological saline of 9% concentration.
Further, person joint's liquid mescenchymal stem cell excretion body extract solution is the concentrate via centrifugation purification.
Further, person joint's liquid mesenchyma used by person joint's liquid mescenchymal stem cell excretion body extract solution is dry
Cell is the third generation to band six generations stem cell.
Further, the storage temperature of person joint's liquid mescenchymal stem cell excretion body extract solution is subzero 20 degree.
Treating rheumatic ostealgia disease treatment injection as described above and preparation method thereof, which includes the following steps：
S1：Person joint's liquid mescenchymal stem cell is obtained, person joint's liquid of acquisition is launched in sterile centrifugation tube, and is added
PBS buffer solution is uniformly mixed, and is then centrifuged with cell Separating tube and is obtained cell separating liquid, supernatant liquor is removed, and will
Lower sediment thing is added in PBS buffer solution and washs, and person joint's liquid mescenchymal stem cell is obtained after centrifuging again；
S2：Person joint's liquid mescenchymal stem cell is cultivated, person joint's liquid mescenchymal stem cell of acquisition is put into culture dish
Culture, person joint's liquid mescenchymal stem cell of hair tonic is the first joint fluid mescenchymal stem cell in culture dish after culture,
By first generation person joint's liquid mescenchymal stem cell of acquisition obtained after five secondary cultures the second generation, the third generation, forth generation,
5th generation, the 6th generation person joint's liquid mescenchymal stem cell；
S3：The rush of person joint's liquid mescenchymal stem cell excretion body point selects the 6th generation person joint's liquid mescenchymal stem cell to set
It in the incubator that oxygen content is 25%, is cultivated using high speed culture medium, when cell bout degree is 75%, uses promotion instead
Excretion body is synthesized carries out culture 7 days with the culture medium of secretion；
S4：Person joint's liquid mescenchymal stem cell excretion body extracts, and will promote the culture medium of excretion body synthesis and secretion in S3
After carrying out culture 7 days, standing collects supernatant liquor i.e. person joint's liquid mescenchymal stem cell excretion body, then passes through the clear liquid of collection
The concentrate for obtaining person joint's liquid mescenchymal stem cell excretion body extract after centrifugation purifies again is crossed, concentrate is statically placed in zero
Lower 20 degree are sealed up for safekeeping；
S5：Synthesize injection, take the concentrate of person joint's liquid mescenchymal stem cell excretion body extract of 20% concentration with
The physiological saline of 9% concentration obtains injection after being thoroughly mixed.
By carrying out composition detection to four groups of embodiments, and one group for the treatment of rheumatic ostealgia disease treatment note is picked from the market
It penetrates agent to be compared, testing result is as follows：
From the experimental data of upper table it is found that the therapeutic effect of the 4th group for the treatment of rheumatic ostealgia disease treatment injection of this programme most
It is good, therefore there is preferable promotional value.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding without departing from the principles and spirit of the present invention can carry out these embodiments a variety of variations, modification, replace
And modification, the scope of the present invention is defined by the appended.
Claims (9)
1. a kind for the treatment of rheumatic ostealgia disease treatment injection with person joint's liquid mescenchymal stem cell excretion body extract, feature
It is；A kind for the treatment of rheumatic ostealgia disease treatment injection with person joint's liquid mescenchymal stem cell excretion body extract includes
Following component：Person joint's liquid mescenchymal stem cell excretion of 5%-20% concentration containing 500 μ l-800 μ l in per 1ml injections
Body extract solution, remaining is the physiological saline of 5% concentration of glucose and 8.5%-9% concentration.
2. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：Person joint's liquid mescenchymal stem cell excretion body extract solution is to be carried via centrifugation
Pure concentrate.
3. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：People used by person joint's liquid mescenchymal stem cell excretion body extract solution is closed
Liquid mescenchymal stem cell is saved as the third generation to band six generations stem cell.
4. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：The storage temperature of person joint's liquid mescenchymal stem cell excretion body extract solution is
Subzero 20 degree.
5. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：A kind of rheumatic bone with person joint's liquid mescenchymal stem cell excretion body extract
Pain disease treatment injection includes following component：Person joint's liquid mesenchyma containing 500 μ l, 5% concentration is dry in per 1ml injections
Cell excretion body extract solution, remaining is the glucose of 5% concentration.
6. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：A kind of rheumatic bone with person joint's liquid mescenchymal stem cell excretion body extract
Pain disease treatment injection includes following component：Person joint's liquid mesenchyma containing 600 μ l, 10% concentration in per 1ml injections
Stem cell excretion body extract solution, remaining is the physiological saline of 8.5% concentration.
7. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：A kind of rheumatic bone with person joint's liquid mescenchymal stem cell excretion body extract
Pain disease treatment injection includes following component：Person joint's liquid mesenchyma containing 700 μ l, 15% concentration in per 1ml injections
Stem cell excretion body extract solution, remaining is the physiological saline of 9% concentration.
8. a kind for the treatment of rheumatic ostealgia disease with person joint's liquid mescenchymal stem cell excretion body extract according to claim 1
Disease treatment injection, it is characterised in that：A kind of rheumatic bone with person joint's liquid mescenchymal stem cell excretion body extract
Pain disease treatment injection includes following component：Person joint's liquid mesenchyma containing 800 μ l, 20% concentration in per 1ml injections
Stem cell excretion body extract solution, remaining is the physiological saline of 9% concentration.
9. described as described in any one of claim 1-8 a kind of carrying person joint's liquid mescenchymal stem cell excretion body extract
Treating rheumatic ostealgia disease treatment injection preparation method, it is characterised in that：The preparation method includes the following steps：
S1：Person joint's liquid mescenchymal stem cell is obtained, person joint's liquid of acquisition is launched in sterile centrifugation tube, and PBS is added
Buffer solution is uniformly mixed, and is then centrifuged with cell Separating tube and is obtained cell separating liquid, supernatant liquor removed, and by lower layer
Sediment is added in PBS buffer solution and washs, and person joint's liquid mescenchymal stem cell is obtained after centrifuging again；
S2：Person joint's liquid mescenchymal stem cell is cultivated, person joint's liquid mescenchymal stem cell of acquisition is put into culture dish and is cultivated,
Person joint's liquid mescenchymal stem cell of hair tonic is first generation person joint's liquid mescenchymal stem cell in culture dish after culture, will be obtained
The first generation person joint's liquid mescenchymal stem cell obtained obtains the second generation, the third generation, forth generation, the 5th after five secondary cultures
Generation, the 6th generation person joint's liquid mescenchymal stem cell；
S3：The rush of person joint's liquid mescenchymal stem cell excretion body point selects the third generation dry thin to the 6th generation person joint's liquid mesenchyma
Born of the same parents are placed in the incubator that oxygen content is 25%, are cultivated using high speed culture medium, when cell bout degree is 75%, use instead
The synthesis of excretion body and the culture medium of secretion is promoted to carry out culture 3-7 days；
S4：Person joint's liquid mescenchymal stem cell excretion body extracts, and the synthesis of excretion body and the culture medium of secretion will be promoted to carry out in S3
After culture 3-7 days, standing collects supernatant liquor i.e. person joint's liquid mescenchymal stem cell excretion body, then passes through the clear liquid of collection
The concentrate for obtaining person joint's liquid mescenchymal stem cell excretion body extract after centrifugation purification again, concentrate is statically placed in subzero
20 degree are sealed up for safekeeping；
S5：Injection is synthesized, the concentrate point of person joint's liquid mescenchymal stem cell excretion body extract of 5%-20% concentration is taken
Injection is obtained after not being thoroughly mixed with the glucose of the physiological saline of 8.5%-9% concentration and 5% concentration.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109985064A (en) * | 2019-05-16 | 2019-07-09 | 北京京蒙细胞生物科技股份有限公司 | Mescenchymal stem cell secretes the purposes of extract, mescenchymal stem cell secretion extract and preparation method thereof |
| CN111484978A (en) * | 2019-01-28 | 2020-08-04 | 深圳市第二人民医院 | miR-140-5p overexpression modified human umbilical cord mesenchymal stem cell, and treatment preparation, preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103917275A (en) * | 2011-07-28 | 2014-07-09 | 耶稣圣婴儿童医院 | Microvesicles isolated from mesenchymal stem cells for use as immunosuppressive agents |
| CN104771413A (en) * | 2014-12-09 | 2015-07-15 | 周治宇 | Application of arthroscopic flushing fluid sourced mesenchymal stem cells in regenerative repair |
| CN105687244A (en) * | 2016-02-02 | 2016-06-22 | 广州赛莱拉干细胞科技股份有限公司 | Preparation, as well as preparation method and application thereof |
| CN107519207A (en) * | 2017-08-31 | 2017-12-29 | 广东颜值科技有限公司 | A kind of immunosuppressant cell preparation and its preparation method and application |
| US20180036348A1 (en) * | 2016-08-04 | 2018-02-08 | Aidan Research & Consulting LLC | Intrathecal administration of mesenchymal stem cells and derivatives thereof for treatment of pain |
| WO2018071677A1 (en) * | 2016-10-13 | 2018-04-19 | Vbc Holdings Llc | Medical uses of exosomes |
-
2018
- 2018-07-20 CN CN201810803937.0A patent/CN108542917A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103917275A (en) * | 2011-07-28 | 2014-07-09 | 耶稣圣婴儿童医院 | Microvesicles isolated from mesenchymal stem cells for use as immunosuppressive agents |
| CN104771413A (en) * | 2014-12-09 | 2015-07-15 | 周治宇 | Application of arthroscopic flushing fluid sourced mesenchymal stem cells in regenerative repair |
| CN105687244A (en) * | 2016-02-02 | 2016-06-22 | 广州赛莱拉干细胞科技股份有限公司 | Preparation, as well as preparation method and application thereof |
| US20180036348A1 (en) * | 2016-08-04 | 2018-02-08 | Aidan Research & Consulting LLC | Intrathecal administration of mesenchymal stem cells and derivatives thereof for treatment of pain |
| WO2018071677A1 (en) * | 2016-10-13 | 2018-04-19 | Vbc Holdings Llc | Medical uses of exosomes |
| CN107519207A (en) * | 2017-08-31 | 2017-12-29 | 广东颜值科技有限公司 | A kind of immunosuppressant cell preparation and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| YU ZHU, ET AL.: "Comparison of exosomes secreted by induced pluripotent stem cell-derived mesenchymal stem cells and synovial membrane-derived mesenchymal stem cells for the treatment of osteoarthritis" * |
| 尼玛才让;: "卡加藏药沐浴、足浴散治疗风湿骨痛临床观察" * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111484978A (en) * | 2019-01-28 | 2020-08-04 | 深圳市第二人民医院 | miR-140-5p overexpression modified human umbilical cord mesenchymal stem cell, and treatment preparation, preparation method and application thereof |
| CN109985064A (en) * | 2019-05-16 | 2019-07-09 | 北京京蒙细胞生物科技股份有限公司 | Mescenchymal stem cell secretes the purposes of extract, mescenchymal stem cell secretion extract and preparation method thereof |
| CN109985064B (en) * | 2019-05-16 | 2020-12-04 | 北京京蒙细胞生物科技股份有限公司 | Application of mesenchymal stem cell secretory extract, mesenchymal stem cell secretory extract and preparation method thereof |
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