CN105326864A - Stem cell-based preparation for treating systemic lupus erythematosus and preparation method thereof - Google Patents
Stem cell-based preparation for treating systemic lupus erythematosus and preparation method thereof Download PDFInfo
- Publication number
- CN105326864A CN105326864A CN201510850440.0A CN201510850440A CN105326864A CN 105326864 A CN105326864 A CN 105326864A CN 201510850440 A CN201510850440 A CN 201510850440A CN 105326864 A CN105326864 A CN 105326864A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- culture
- cell
- medicine
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a stem cell-based preparation for treating systemic lupus erythematosus and a preparation method thereof. The preparation is prepared from mesenchymal stem cells, epidermal growth factors, human albumin, interleukin 11 and normal saline according to a certain proportion. The preparation method disclosed by the invention has the advantages of rich resource, convenience in taking materials, simplicity in manufacturing and low immunological rejection after cell transplantation; according to the stem cell-based preparation and the preparation method, the mesenchymal stem cells are combined with cell factors for the first time, so that the treatment effect is obvious, and a method for treating the systemic lupus erythematosus is basically provided.
Description
Technical field
The invention belongs to biotechnology and regenerative medicine field, be specifically related to stem cell medicine of a kind of systemic lupus erythematosus and preparation method thereof.
Background technology
Systemic lupus erythematosus (sle) (systemiclupuserythematosus; SLE) be a kind of autoimmune disease of involving multisystem multiple organ; immune system dysfunction in patient body; T, bone-marrow-derived lymphocyte abnormal activation; produce multiple autoantibody and immune complex, cause the multiple organs of whole body such as patient skin, joint, kidney and system to suffer damage.SLE is apt to occur in young women, M-F is 1:9, in China adult female, sickness rate is 0.1%, higher than Hesperian 0.05%, and along with the increase of socioeconomic development and life stress, the sickness rate of SLE rises day by day, because this course of disease is grown and recurrent exerbation, brings great impact to the psychology of patient and physiology.
The conventional Therapeutic Method of SLE, mainly by taking the medicine such as glucocorticoid or immunosuppressant, alleviates the effect of inflammatory reaction because immune system attack normal structure causes and Immunosuppression system with this.Although glucocorticoid and immunosuppressant are used to the systemic lupus erythematosus obvious improvement that made the prognosis of patient have, the current treatment status of SLE still allows of no optimist: 1. still have an appointment at present 10% patient dead in morbidity 5 years; 2. due to the side effect that progress and some treatments of disease bring, as there are infection, bone marrow depression, osteoporosis, hypertension, diabetes etc., the especially application of cell toxicity medicament and glucocorticoid, the average life of SLE patient is significantly shorter than normal population; 3. conventional Therapeutic Method can only slow down the development speed of the state of an illness, non-radical curing of disease.Therefore, find that some are efficient, the newly side of controlling of few side effects is extremely urgent.
Since within 1996, starting to move with autologous stem cell (HSC) and growing treatment SLE, good curative effect is achieved in the intractable SLE of serious symptom, but there is the problem of easily recurrence, though and allogene HSC move grow treatment SLE can make some patients state of an illness long-term remission, but donor source is not enough, case fatality rate is up to 30% ~ 50%, and medical expense is high.Except HSC, also there is similar fibroblastic mescenchymal stem cell (MSC) in a kind of form in bone marrow, MSC have multi-lineage potential, immunogenicity low with growth rate the feature such as fast.MSC not only has the function of hematopoiesis support, also there is the function of immunomodulating and inducing immune tolerance, research shows that MSC plays the mechanism of immunomodulating and inducing immune tolerance mainly: suppressor T cell is bred, the cell number of generation interleukin-4 (IL-4) is reduced, the cell number producing interferon gamma (IFN-γ) increases, and regulates the balance of Th1/Th2.MSC regulates the release of inflammatory factor, inducing immune tolerance, suppression of autoimmune responses thus reach and treat SLE long-term effectively by number of ways.But because in simple bone marrow stem cell colony, the content of MSC is few, if not by external further amplification, be difficult to reach therapeutic effect, and SLE is a kind of systemic disease, simple use MSC treats, therapeutic effect is limited, particularly for some serious symptom SLE patients, is difficult in a short time reach good therapeutic effect.
In order to solve the problem, the invention provides a kind of stem cell medicine being used for the treatment of SLE and preparation method thereof, stem-cell therapy SLE be had to the clinical meaning of particular importance.
Summary of the invention
At present, the main Problems existing of the Therapeutic Method of SLE and defect: 1. conventional treatments side effect is large, relapse rate is high, therapeutic effect is not good, failing to respond to any medical treatment to intractable SLE; 2. stem cell moves and grows as novel therapies, and autologous HSC moves that to grow relapse rate high; Allosome HSC move grow distribution type difficulty, mortality rate is high; 3. adult MSC moves that to grow be current the most effective Therapeutic Method, and its immunogenicity is low, but limited amount, and belongs to systemic disease due to SLE, and simple employing MSC treats, and therapeutic effect is limited.In order to solve the problem and defect, the object of this invention is to provide a kind of stem cell medicine for the treatment of SLE and preparation method thereof, make up the weak point existed in the current Therapeutic Method of SLE.
The invention provides and be a kind ofly used for the treatment of stem cell preparation method in the stem cell medicine of systemic lupus erythematosus (sle), the method is with specific process induction and cultivate embryonic stem cell, be prepared into stem cell medicine, can effectively improve and strengthen the curative effect of stem-cell therapy systemic lupus erythematosus (sle).
For realizing above-mentioned technical purpose, reach above-mentioned technique effect, the present invention is achieved through the following technical solutions:
A preparation method for the stem cell medicine of systemic lupus erythematosus, mainly comprises the following steps:
(1) In vitro culture of embryonic stem cell, amplification: after adopting the culture fluid of embryonic stem cell that embryonic stem cell is resuspended, adjustment cell density is (1 ~ 3) × 10
4/ cm
2, be inoculated in the T175 Tissue Culture Flask of gelatin bag quilt, be placed in 37 DEG C, 5%CO
2hatch in incubator, fluid infusion after 48 hours, after 72 hours, change liquid.Until cell fusion to 85 ~ 90% time with 0.25% trypsinization go down to posterity, go down to posterity in 100mm culture dish according to 1:3 ~ 1:5, supply culture fluid continue cultivate, every 2 ~ 3d Digestive system had digestive transfer culture;
(2) inducing embryo stem cell to Derived from Mesenchymal Stem Cells, cultivate, go down to posterity and detect: the cell concentration of gained embryonic stem cell in step (1) is adjusted to 5 × 10
5/ ware is inoculated in 100mm culture dish, and every ware adds induction system 8ml, culture dish is placed in 37 DEG C, 5%CO
2carry out inducing culture 24h; Stem cell after inducing and giving birth to is changed culture fluid, and adopt Mesenchymal stem cell nutrient solution, go down to posterity with 0.25% trypsinization after cell covers with, amplification culture to stem cell number is 10
8-10
9; With flow cytometer, the mescenchymal stem cell after P3 generation induction is identified: described mescenchymal stem cell expresses CD73, CD90, CD105 and CD29, does not express CD34 and CD14;
(3) mescenchymal stem cell step (2) prepared and epidermal growth factor (EGF), interleukin 11 (IL-11), human albumin mix in proportion, and surplus normal saline is supplied, and is prepared into stem cell medicine;
Wherein, consisting of of Embryonic Stem Cell media in described step (1): 90%DMEM culture medium, 10% hyclone, 0.1mmol/L beta-mercaptoethanol, 1mmol/LL-glutamine, 1% non essential amino acid, 4ng/ml recombination human basic fibroblast somatomedin;
Induction system in described step (2) is: containing DMEM (high sugar) culture medium of removal LIF, 2mmol/LL-glutamine of 10% hyclone, 100ng/mLActivinA, 10 μm of ol/LY-27632 (Rho inhibitors of kinases);
Consisting of of the middle Mesenchymal stem cell nutrient solution of described step (2): 15% hyclone (FBS), 85%DMEM culture fluid;
Specifically consisting of of the middle stem cell medicine of described step (3): the concentration of mescenchymal stem cell is (4 ~ 6) × 10
7individual/ml, EGF is 8ng/mL, and the mass volume ratio of human albumin is 2%, IL-11 is 30ng/mL, and surplus is normal saline.
A kind of stem cell medicine of systemic lupus erythematosus is prepared from by above-mentioned steps, wherein formulated by a certain percentage by the mescenchymal stem cell after inducing, epidermal growth factor (EGF), human albumin, interleukin 11 (IL-11), normal saline;
Wherein, specifically the consisting of of described stem cell medicine: the concentration of mescenchymal stem cell is (4 ~ 6) × 10
7individual/ml, EGF is 8ng/mL, and the mass volume ratio of human albumin is 2%, IL-11 is 30ng/mL, and surplus is normal saline.
Further, described mescenchymal stem cell is by induced differentiation of embryonic stem cells.
Further, described embryonic stem cell is the class cell separated in body early embryo (before gastrula stage) or original gonad, and it has the characteristic of In vitro culture infinite multiplication, self renewal and Multidirectional Differentiation.
The present invention has the following advantages and effect relative to prior art:
(1) the present invention uses by human embryo stem cell external evoked mesenchyma stem cell combined cytokine of turning out first, and carry out the treatment of SLE, therapeutic effect is remarkable, fundamentally provides a kind of method for the treatment of SLE.
(2) the present invention makes up by mesenchyma stem cell combined cytokine being used the weak point existed in the current Therapeutic Method of SLE, wherein: 1. mescenchymal stem cell mainly has the advantage of the following aspects: a multi-lineage potential, can differentiate multiple normal cell and repair body damaged tissues; B immunogenicity is low, greatly reduces the probability that graft versus host disease occurs; C growth rate is fast, and preparation easily, can large-scale industrialized production; D has the function of immunomodulating and inducing immune tolerance, and suppressor T cell is bred, and regulates the balance of Th1/Th2, suppression of autoimmune responses thus reach and treat SLE long-term effectively.2. the existence energy inducing mesenchymal stem cell of the advantage of EGF: EGF breaks up to skin progenitor cell, patient SLE of 75% shows the skin damage phenomenon such as cutin desquamation and hair follicle bolt, oral ulcer, photosensitivity clinical, skin progenitor cell can migration and differentiation be stratum basale downwards on the one hand, and then generates hair follicle; Then upwards can move on the other hand, be divided into various epidermis cell, and then repair the damaged skin of SLE patient.3. the advantage of IL-11: thrombocytopenia appears in 50%SLE patient, IL-11 can reduce the hemorrhage and death that SLE patient patient causes because of thrombocytopenia.
(3) after in stem cell medicine input SLE patient body provided by the invention, cell energy long-term surviving is in patient body, serve permanently effective therapeutical effect, when avoiding employing conventional treatments, the shortcoming of the various immunosuppressant of needs of patients long-term taking and hormone medicine.
Above-mentioned explanation is only the general introduction of technical solution of the present invention, in order to better understand technological means of the present invention, and can be implemented according to the content of description, coordinates accompanying drawing to be described in detail as follows below with better case study on implementation of the present invention.The specific embodiment of the present invention is provided in detail by following examples and accompanying drawing thereof.
Accompanying drawing explanation
Accompanying drawing described herein is used to provide a further understanding of the present invention, and form a application's part, schematic description and description of the present invention, for explaining the present invention, does not form inappropriate limitation of the present invention.In the accompanying drawings:
Fig. 1 is growth of mesenchymal stem cells curve chart, and abscissa is cultivated days, and vertical coordinate is absorbance;
Fig. 2 is 24 urine protein quantitation detection figure of lupus mice, respectively illustrates three groups of mices A1, A2, A3 with advancing age, the situation of change of urine protein in figure, and at the age of mice when abscissa is detection, vertical coordinate is urine protein content;
Fig. 3 is the body weight growth record of lupus mice, respectively illustrates three groups of mices A1, A2, A3 with advancing age, the situation of change of body weight in figure.At the age of mice when abscissa is detection, vertical coordinate is body weight;
Fig. 4 is three groups of mices A1, A2, A3 anti-double-chain DNA (dsDNA) antibody titer mensuration figure at pre-treatment and after treatment.
Detailed description of the invention
Below with reference to the accompanying drawings and the present invention is described in further detail in conjunction with the embodiments, but embodiments of the present invention are not limited thereto.
The In vitro culture of embodiment 1 embryonic stem cell:
By the embryonic stem cell preserved in liquid nitrogen in 37 DEG C of water-baths, flash melt, cell suspension becomes liquid state from solid-state.After adopting DMEM culture medium that embryonic stem cell is resuspended, adjustment cell density is (1 ~ 3) × 10
4/ cm
2, be inoculated in the T175 Tissue Culture Flask of gelatin bag quilt, be placed in 37 DEG C, 5%CO
2hatch in incubator, fluid infusion after 48 hours, after 72 hours, change liquid.Until cell fusion to 80 ~ 90% time %0.25 trypsinization go down to posterity, go down to posterity in 100mm culture dish according to 1:3 ~ 1:5, supply culture fluid continue cultivate, every 2 ~ 3d Digestive system had digestive transfer culture.
Embodiment 2 inducing embryo stem cell is to Derived from Mesenchymal Stem Cells
In vitro culture obtains the method for embryonic stem cell, with embodiment 1
Inducing embryo stem cell is to Derived from Mesenchymal Stem Cells: the cell concentration of gained embryonic stem cell suspension in step (1) is adjusted to 5 × 10
5/ ware is inoculated in 100mm culture dish, and every ware adds induction system 8ml, culture dish is placed in 37 DEG C, 5%CO
2carry out inducing culture 24h.Described induction system is: containing DMEM (high sugar) culture medium of removal LIF, 2mmol/LL-glutamine of 10% hyclone, 100ng/mLActivinA, 10 μm of ol/LY-27632 (Rho inhibitors of kinases).
The cultivation of embodiment 3 mescenchymal stem cell, go down to posterity and detect
Obtain the method for mescenchymal stem cell, with embodiment 1,2
The cultivation of mescenchymal stem cell: change culture fluid, adopts Mesenchymal stem cell nutrient solution (consisting of: 15% hyclone, the DMEM culture fluid of 85%), is positioned over 37 DEG C of CO2 gas incubator and cultivates.Cell culture 3-4d, changes a not good liquor in every 2 days, treats that cell fusion degree reaches 85%-90%, abandon supernatant, with 0.25% trypsinization results P1 for mescenchymal stem cell.By gathered in the crops cell according to 5 × 10
3/ cm
2be inoculated in 100mm culture dish and be positioned over 37 DEG C of CO2 gas incubator cultivations, cultivate 2-3d, cell fusion degree reaches 85%-90%, with the trypsinized liquid digestion harvesting of 0.25%.Every 2-3d had digestive transfer culture, amplification culture to 10
8-10
9, by for subsequent use for the cell harvesting being cultured to for 3rd ~ 5 generations.
Mescenchymal stem cell surface marker Testing and appraisal: digestion collection the 3rd generation mesenchymal cell, the centrifugal 5min of 1500rpm; Abandon supernatant, add D-Hank
,s solution is resuspended, draws 100 μ l cell suspension (about 2.3 × 10
5individual cell) add in fluidic cell pipe; Respectively to CDl05, CD90, CD29, CD73, CDl4, CD34 humanized murine antibodies adding 10 μ lPE labellings in each pipe, negative control pipe adds Mus lgG-PE; 4 DEG C, lucifuge reaction 30min, PBS wash once, the centrifugal 5min of 1500rpm; Abandon supernatant, add 200 μ lPBS blow and beat mixing cells, 200 μ l1% paraformaldehydes are fixed, put 4 DEG C to be measured, in 3 days flow cytometer detect.Result surface cultured cells through FCM analysis, negative control: 0.18%; Positive expression: CD105-97.32%, CD105-97.32%, CD90-98.55%, CD29-93.17%, CD73-92.84%; And negative expression: CD14-1.38%, CD34-0.79%.
Growth of mesenchymal stem cells determination of activity: collection the 3rd generation mescenchymal stem cell, adjustment cell concentration 1 × 10
4individual/mL is inoculated in 96 porocyte culture plates, 7 groups, often organizes 5 holes, every hole 100 μ L, 37 DEG C of incubator quiescent culture; Within continuous 7 days, measure absorbance with mtt assay after inoculation, in test group, in every hole, add the 0.5%MTT(5mg/mL with PBS preparation) 20uL, cultivates in 37 DEG C of incubators; Stop after 4h cultivating, by careful for culture fluid in hole sucking-off, every hole adds 150 μ L dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.Often row first hole is set to zeroing hole (culture medium, MTT, dimethyl sulfoxide) simultaneously.Measure each hole light absorption value with microplate reader OD490nm place, get average and draw formation curve.As shown in Figure 1, the 1st ~ 2d poor growth after mescenchymal stem cell goes down to posterity, is in lag phase to result, and from 3d, Growth of Cells is rapid, and enter exponential phase, 5d peaks; Afterwards, Growth of Cells is slow, enters plateau.
The preparation method of embodiment 4 stem cell medicine
Collect through detecting and the cell of assay approval, and detect cell viability, living cells is more than 95%, and cell number reaches the requirement of cell preparation: mescenchymal stem cell number is 5.2 × 10
7individual/ml, abandons supernatant after cell suspension is centrifugal, and be resuspended in containing 8ng/mL epidermal growth factor, 2% human albumin, 30ng/mLIL-11 1ml normal saline in, be prepared into stem cell medicine.
Embodiment 5 stem cell medicine treatment lupus mice animal model experiment
Lupus mice stem cell medicine injection experiment: to choose 30 body weight be 21 ± 3g age is 6-7 week female mice, be divided into three groups at random, blank group (A1, n=10), only carry out the treatment group (A2 of mescenchymal stem cell injection, n=10), the treatment group (A3, n=10) of stem cell medicine of the present invention injection is adopted.A2 adopted the mode of tail vein injection to inject 3 × 10 in the 17th, 18,19 week
7individual third generation mescenchymal stem cell, A3 adopted the mode of tail vein injection to inject 3 × 10 in the 17th, 18,19 week
7individual third generation stem cell medicine of the present invention, A1 is injected 0.5ml normal saline.
(1) impact of stem cell medicine injection on lupus mice 24h urine protein quantitation
Within every 3 weeks from the 16th week, measure mice 24h excretion quantity of urinary protein, adopt metabolic cage to leave and take mice twenty-four-hour urine amount, and measure its protein content with Coomassie Brilliant Blue.Measurement result is as shown in Figure 2: blank group A1 was from 16 weeks age, and urine protein raises gradually, and treatment group A2 and A3 is at the treatment initial stage, and urine protein also rises gradually, but more blank group of ascendant trend slowly, and after 22 weeks, treatment group urine protein level tends towards stability; To treatment group A3 urine protein level (1.65 scholars l.13mg) when the 31st week significantly lower than A2 (2.52 scholar 1.26mg) (p<0.05) and blank group A1 (5.62 scholar 2.14mg) (p<0.01).
(2) impact of stem cell medicine injection on lupus mice routine blood test, serum creatinine
After lupus mice being carried out to the injection of stem cell medicine, when mice grows to the 31st week, carry out routine blood test to A1, A2, A3 group mice respectively, the mensuration of serum creatinine, wherein routine blood test comprises: leukocyte, serum albumin, platelet.Measurement result is as shown in table 1, along with the development of the state of an illness, and blank group A1 leukocyte (2.13 soil 0.82 × 10
9/ L), serum albumin (19 ± 4g/L), platelet (74.76 soil 14.55 × 10
9/ L) all lower than normal value, each index for the treatment of group A2 routine blood test rises all to some extent, but platelet and serum albumin are appointed so lower than normal value, and arriving of the treatment group A3 state of an illness controls, routine blood test recovers normal; The serum creatinine measured value of three groups of mices is A2(10.2 scholar 2.1 μm of ol/L) and A3(6.5 scholar 1.7 μm of ol/L) all remarkable in blank group A1(29.5 soil 3.8 μm of ol/L) (p<0.01).
The impact of table 1 stem cell medicine injection on lupus mice routine blood test, serum creatinine
(3) impact of stem cell medicine injection on lupus mice body weight
From the 16th week, within every 4 weeks, carry out body weight determination respectively to A1, A2, A3 group mice, measurement result as shown in Figure 3.When just starting, blank group A1 body weight slowly rises, but body weight decreases after 24 weeks; Treatment group A2, A3 body weight rise very fast, tend towards stability after 24 weeks, still on the rise; Treat each group compared with blank group difference have statistical significance, and A2, A3 compare without significant difference.
(4) impact of stem cell medicine injection on lupus mice anti-double-chain DNA (dsDNA) antibody titer
Before carrying out stem cell medicine treatment injection, first three groups of mices of the 16th week are carried out to the mensuration of anti-double-chain DNA (dsDNA) antibody titer, after stem cell medicine injection is carried out to A2, A3 group, when mice grows to the 31st week, again three groups of mices are carried out to the mensuration of anti-double-chain DNA (dsDNA) antibody titer.Measurement result as shown in Figure 4, before carrying out stem cell medicine injection, three groups of little mouse-anti (dsDNA) antibody titers are suitable, after injection, anti-(dsDNA) antibody titer for the treatment of group A3 is that (36.54 scholar 15.28IU/ml) is significantly lower than A2(66.16 scholar 26.12IU/ml) (p<0.01) and blank group A1(87.26 scholar 32.16IU/ml) (p<0.01).
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (4)
1. the stem cell medicine of a systemic lupus erythematosus, it is characterized in that, mainly comprise the following steps: the In vitro culture of (1) embryonic stem cell, amplification: after adopting the culture fluid of embryonic stem cell that embryonic stem cell is resuspended, adjustment cell density is (1 ~ 3) × 10
4/ cm
2, be inoculated in the T175 Tissue Culture Flask of gelatin bag quilt, be placed in 37 DEG C, 5%CO
2hatch in incubator, fluid infusion after 48 hours, after 72 hours, change liquid; Until cell fusion to 85 ~ 90% time with 0.25% trypsinization go down to posterity, go down to posterity in 100mm culture dish according to 1:3 ~ 1:5, supply culture fluid continue cultivate, every 2 ~ 3d Digestive system had digestive transfer culture;
(2) inducing embryo stem cell to Derived from Mesenchymal Stem Cells, cultivate, go down to posterity and detect: the cell concentration of gained embryonic stem cell in step (1) is adjusted to 5 × 10
5/ ware is inoculated in 100mm culture dish, and every ware adds induction system 8ml, culture dish is placed in 37 DEG C, 5%CO
2carry out inducing culture 24h; Stem cell after inducing and giving birth to is changed culture fluid, and adopt Mesenchymal stem cell nutrient solution, go down to posterity with 0.25% trypsinization after cell covers with, amplification culture to stem cell number is 10
8-10
9; With flow cytometer, the mescenchymal stem cell after P3 generation induction is identified: described mescenchymal stem cell expresses CD73, CD90, CD105 and CD29, does not express CD34 and CD14;
(3) mescenchymal stem cell step (2) prepared and epidermal growth factor (EGF), interleukin 11 (IL-11), human albumin mix in proportion, and surplus normal saline is supplied, and is prepared into stem cell medicine;
Wherein, consisting of of Embryonic Stem Cell media in described step (1): 90%DMEM culture medium, 10% hyclone, 0.1mmol/L beta-mercaptoethanol, 1mmol/LL-glutamine, 1% non essential amino acid, 4ng/ml recombination human basic fibroblast somatomedin;
Induction system in described step (2) is: containing DMEM (high sugar) culture medium of removal LIF, 2mmol/LL-glutamine of 10% hyclone, 100ng/mLActivinA, 10 μm of ol/LY-27632 (Rho inhibitors of kinases);
Consisting of of the middle Mesenchymal stem cell nutrient solution of described step (2): 15% hyclone (FBS), 85%DMEM culture fluid;
Specifically consisting of of the middle stem cell medicine of described step (3): the concentration of mescenchymal stem cell is (4 ~ 6) × 10
7individual/ml, EGF is 8ng/mL, and the mass volume ratio of human albumin is 2%, IL-11 is 30ng/mL, and surplus is normal saline.
2. the stem cell medicine of the systemic lupus erythematosus be prepared from by step described in claim 1, it is characterized in that, formulated by a certain percentage by the mescenchymal stem cell after inducing, epidermal growth factor (EGF), human albumin, interleukin 11 (IL-11), normal saline;
Wherein, specifically the consisting of of described stem cell medicine: the concentration of mescenchymal stem cell is (4 ~ 6) × 10
7individual/ml, EGF is 8ng/mL, and the mass volume ratio of human albumin is 2%, IL-11 is 30ng/mL, and surplus is normal saline.
3. the stem cell medicine of a kind of systemic lupus erythematosus according to claim 2, is characterized in that, described mescenchymal stem cell is by induced differentiation of embryonic stem cells.
4. the stem cell medicine of a kind of systemic lupus erythematosus according to claim 2, it is characterized in that, described embryonic stem cell is the class cell separated in body early embryo (before gastrula stage) or original gonad, and it has the characteristic of In vitro culture infinite multiplication, self renewal and Multidirectional Differentiation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510850440.0A CN105326864A (en) | 2015-11-30 | 2015-11-30 | Stem cell-based preparation for treating systemic lupus erythematosus and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510850440.0A CN105326864A (en) | 2015-11-30 | 2015-11-30 | Stem cell-based preparation for treating systemic lupus erythematosus and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105326864A true CN105326864A (en) | 2016-02-17 |
Family
ID=55277770
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510850440.0A Pending CN105326864A (en) | 2015-11-30 | 2015-11-30 | Stem cell-based preparation for treating systemic lupus erythematosus and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105326864A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111420034A (en) * | 2019-01-08 | 2020-07-17 | 上海莱馥医疗科技有限公司 | Mixed stem cell preparation for treating psoriasis and preparation method thereof |
| CN116218771A (en) * | 2022-12-30 | 2023-06-06 | 广东医科大学附属医院 | hUC-MSCs preparation and preparation method and application thereof |
| WO2024041162A1 (en) * | 2022-08-24 | 2024-02-29 | 深圳市北科生物科技有限公司 | Mesenchymal stem cell and exosome thereof obtained by treatment of at least two cytokines of il4, il21, and il27, and use thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013082543A1 (en) * | 2011-11-30 | 2013-06-06 | Advanced Cell Technology, Inc. | Mesenchymal stromal cells and uses related thereto |
| CN104487568A (en) * | 2012-07-11 | 2015-04-01 | 爱姆斯坦生物技术公司 | Mesenchymal-like stem cells derived from human embryonic stem cells, methods and uses thereof |
-
2015
- 2015-11-30 CN CN201510850440.0A patent/CN105326864A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013082543A1 (en) * | 2011-11-30 | 2013-06-06 | Advanced Cell Technology, Inc. | Mesenchymal stromal cells and uses related thereto |
| CN104487568A (en) * | 2012-07-11 | 2015-04-01 | 爱姆斯坦生物技术公司 | Mesenchymal-like stem cells derived from human embryonic stem cells, methods and uses thereof |
Non-Patent Citations (11)
| Title |
|---|
| BENJAMIN E. REUBINOFF ET AL.: "Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro", 《NATURE BIOTECHNOLOGY》 * |
| YONGXING LIU ET AL.: "One-Step Derivation of Mesenchymal Stem Cell (MSC)-Like Cells from Human Pluripotent Stem Cells on a Fibrillar Collagen Coating", 《PLOS ONE》 * |
| 刘志超等: "《细胞因子与临床疾病》", 31 August 2008, 海南出版社 * |
| 吴蓉蓉: "人胚胎干细胞建系，鉴定及分化为多能性间充质样前体细胞", 《中国博士学位论文全文数据库-医药卫生科技辑》 * |
| 徐如祥: "《神经干细胞》", 31 December 2006, 军事医学科学出版社 * |
| 杨桂鲜: "间充质干细胞移植治疗系统性红斑狼疮研究进展", 《国际泌尿系统杂志》 * |
| 石伦刚等: "人胚胎干细胞向类间充质干细胞的诱导分化及鉴定", 《上海交通大学学报（医学版）》 * |
| 祁秋干等: "脐血间充质干细胞治疗严重性系统性红斑狼疮的临床疗效初探", 《中华细胞与干细胞杂志》 * |
| 罗振: "《生命不老的源泉-干细胞》", 31 July 2014, 吉林人民出版社 * |
| 陆士新: "《干细胞与肿瘤》", 31 July 2009, 中国协和医科大学出版社 * |
| 陈临溪等: "《细胞信号转导药理与临床》", 31 October 2014, 人民军医出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111420034A (en) * | 2019-01-08 | 2020-07-17 | 上海莱馥医疗科技有限公司 | Mixed stem cell preparation for treating psoriasis and preparation method thereof |
| WO2024041162A1 (en) * | 2022-08-24 | 2024-02-29 | 深圳市北科生物科技有限公司 | Mesenchymal stem cell and exosome thereof obtained by treatment of at least two cytokines of il4, il21, and il27, and use thereof |
| CN116218771A (en) * | 2022-12-30 | 2023-06-06 | 广东医科大学附属医院 | hUC-MSCs preparation and preparation method and application thereof |
| CN116218771B (en) * | 2022-12-30 | 2023-12-19 | 广东医科大学附属医院 | hUC-MSCs preparation and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103330720B (en) | Mixing stem cell injection and preparation method thereof | |
| CN103422176B (en) | Construction method of human amniotic mesenchymal stem cell bank | |
| CN104988110A (en) | Method for transforming umbilical cord mesenchymal stem cells into islet cells | |
| CN102643784B (en) | A kind of amplification in vitro system of hematopoietic stem/progenitor | |
| CN103301449B (en) | A kind of preparation method of large-scale culture dendritic cell vaccine and application thereof | |
| CN105779383A (en) | Preparation method and application of adipose-derived stem cell-hydrogel three-dimensional cultivation system | |
| CN102028970A (en) | Stem cell preparation for treating cirrhosis | |
| CN109988746A (en) | A kind of mescenchymal stem cell adipogenic induction differentiation method | |
| CN103497892B (en) | A kind of cell cultures base material and its preparation method and application | |
| CN114181968A (en) | Preparation method and application of artificial blood progenitor cells with B lineage differentiation potential | |
| CN105326864A (en) | Stem cell-based preparation for treating systemic lupus erythematosus and preparation method thereof | |
| CN101831403B (en) | Method for amplifying mesenchymal stem cells of human umbilical cord and placenta in vitro | |
| CN1286970C (en) | Method of prodcing megacaryocyle using umbilical blood CD 344+cell in vitro induction method | |
| CN103421740B (en) | In-vitro culture and proliferation method for human mesenchymal stem cells | |
| CN107287156A (en) | A kind of isolated culture method of fat mesenchymal stem cell and its application | |
| CN1844373A (en) | Method for in vitro preparation of hepatocyte induced by umbilical cord blood hematopoietic stem cell | |
| CN102719397B (en) | Mesenchymal stem cell and application thereof in resisting HIV-1 (human immunodeficiency virus) | |
| CN107080757A (en) | A kind of humanization hepatitis B mouse model of utilization stem cell constructing and application | |
| CN106190946A (en) | A kind of culture medium for expansion of stem cells | |
| CN104988111A (en) | Inducing liquid for converting UC-MSC into islet cells and application thereof | |
| CN102154208A (en) | Preparation method and use of cord blood-derived (CD)133 and brain glioma stem cell antigen carrying dendritic cells | |
| CN104099291A (en) | Method for improving proliferation ability and performance of umbilical cord blood OECs (outgrowth endothelial cells) | |
| CN104419660A (en) | Method for preparing autologous hematopoietic stem cells, kit, the stem cells and application | |
| CN101469322A (en) | Cell preparation for liver injury and hepatic cirrhosis and preparation thereof | |
| CN104726400A (en) | Animal-source-free component culture method for differentiation from human pluripotent stem cells to germ cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160217 |