CN104789530B - It is a kind of to increase the method for Cord blood megakaryoblast directed differentiation quantity - Google Patents
It is a kind of to increase the method for Cord blood megakaryoblast directed differentiation quantity Download PDFInfo
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- CN104789530B CN104789530B CN201510239696.8A CN201510239696A CN104789530B CN 104789530 B CN104789530 B CN 104789530B CN 201510239696 A CN201510239696 A CN 201510239696A CN 104789530 B CN104789530 B CN 104789530B
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Abstract
The present invention discloses a kind of method for improving Cord blood megakaryoblast directed differentiation quantity.This method is oriented differentiation using umbilical cord blood hematopoietic stem cell as sample cell in the defined medium containing RV.The defined medium for containing resveratrol makes megakaryoblast directed differentiation operate more handy and safe and efficient.In umbilical cord blood hematopoietic stem cell into megakaryoblast directed differentiation incubation, RV is added while adding cell factor, the effect of increase megakaryoblast generation quantity has been reached, of the invention is easy to operate, cost is low, and obtained megakaryoblast is safe.
Description
Technical field
The invention belongs to cell technology field, and in particular to one kind increase umbilical cord blood hematopoietic stem cell is fixed to megakaryoblast
To the method for quantity of differentiation.
Background technology
Candidate stem cell has self-renewal capacity and can be divided into the precursor of various haemocytes, ultimately forms blood
Cell, including red blood cell, leucocyte and blood platelet.Candidate stem cell is mainly derived from marrow, Cord blood and peripheral blood, in clinic
It is upper to be used for the various malignant hematologic diseases of transplantation treatment and some immunological diseases.
First case Umbilical Cord Blood Transplantation success in 1989, transplanting/infusion of Cord blood has been clinical treatment blood system so far
The important aspect of disease of uniting and some immunological diseases.As marrow hemopoietic stem cells and peripheral blood hematopoietic stem cells, Cord blood
Before transplantation, receptor will generally carry out Large Dose Irradiation/chemotherapy to reach the effect of clear marrow to candidate stem cell, especially pernicious blood
Liquid tumor patient, after radiotherapy/chemotherapy, the hemopoietic system of receptor is equally also damaged seriously while tumour cell is killed,
The blood platelet famine of receptor within a period of time, causes easy bleeding or even dead, it is necessary to which platelet transfusion is to support.And
It is hematoblastic to recover the major reason that progress is also influence transplanting success or failure after umbilical cord blood stem cell transplantation.It is universal at present
Think that platelet recovery is that megakaryoblast quantity is not abundant in Cord blood and differentiation and maturation is slow more slowly after Umbilical Cord Blood Transplantation
And cause.
Megacaryocyte is that candidate stem cell gradually breaks up and produced, and candidate stem cell is first divided into megakaryoblast
(Megakaryocytic progentitor cell, MKPC) and then the megacaryocyte for being further divided into maturation, is finally released
Release blood platelet.In recent years find, allosome umbilical cord blood hematopoietic stem cell in vitro directed differentiation be megakaryoblast after, in Hematopoietic Stem
It is infused into cell transplantation in recipient's body, is to be expected to solve after radiotherapy/chemotherapy blood platelet after aleukia and Umbilical Cord Blood Transplantation
Recover the important method of delay.Umbilical cord blood hematopoietic stem cell vitro directed differentiation amplification be stem cell field research emphasis it
One, the research broken up to CFU-E, grain system progenitor cells direction more to be had been reported that.Currently, umbilical cord blood hematopoietic stem cell is to huge
The amplification of core progenitor cells directed differentiation is generally carried out using combination of cytokines, generally there is TPO, FLT, IL-3, IL-6, SCF etc..
Resveratrol is a kind of natural plantibody, when plant suffers from the unfavorable conditions such as fungal infection, ultraviolet
The phenolic compound can be produced, protection, defense reaction are risen to plant.Resveratrol is present in Vitaceae, Liliaceae, pulse family etc. 70
In various plants.The anti-cardiovascular disease effect of resveratrol is begun to be recognized by scholars because of " French antinomy ", with world's model
Interior further research is enclosed, the other biological effect of resveratrol is gradually found.Resveratrol has antitumor, anti-inflammatory
Disease, immunological regulation, anti-ageing a variety of physiological actions of waiting for a long time.Found by the research in terms of molecular biology and signalling channel, white lamb's-quarters
Reed alcohol can promote cancer cell-apoptosis by suppressing the signal transduction of MAPK or NF-kB passages.In terms of the differentiation of candidate stem cell,
Resveratrol can accelerate the maturation of CFU-E, and CFU-GM colony formation quantity is improved when expanding in vitro.
Need to remove blood plasma and red blood cell after umbilical cord blood collection, to be enriched with total karyocyte (TNC) rich in candidate stem cell.
Except candidate stem cell, total karyocyte is also containing a large amount of various cells.Complicated composition has bad shadow to amplification in vitro efficiency
Ring.Although can obtain purer candidate stem cell by magnetic bead sorting more to optimize amplification in vitro, increased cost is to needing
Situation about largely expanding has severely restricts, while more operating procedures also bring bigger pollution risk.
The content of the invention
In order to overcome the shortcoming and deficiency of prior art, Cord blood macronucleus ancestral is improved it is an object of the invention to provide one kind
The method of cell directional quantity of differentiation.This method adds natural compound resveratrol, and stem cell products are obtained after culture, makes
Standby process is simple, reliable and reproducible and with low cost.By the above method, the macronucleus required for known clinical practice
During progenitor cell population, it is possible to reduce preparation time, provide more reliable for clinical practice and efficiently support.
The purpose of the present invention is achieved through the following technical solutions：One kind improves Cord blood megakaryoblast directed differentiation quantity
Method, using umbilical cord blood hematopoietic stem cell as sample cell, differentiation is oriented in the defined medium containing RV.
In the described defined medium containing RV the concentration gradient of RV be set in 5~25 μm of ol/L it
Between；More preferably 15 μm ol/L；
The described defined medium containing RV, is added thin in the Stemspan culture mediums of STEMCELL companies
Intracellular cytokine IL-3 (20ng/mL), IL-6 (50ng/mL), SCF (50ng/mL) and TPO (50ng/mL).
A kind of method for improving Cord blood megakaryoblast directed differentiation quantity, including step in detail below：
(1) resveratrol is dissolved in cell-protecting, is configured to resveratrol stock solution；
(2) defined medium is prepared, the resveratrol stock solution that step (1) is prepared is added in defined medium,
It is 0 to obtain resveratrol concentration in the defined medium containing resveratrol, control group；
(3) separation of umbilical cord blood hematopoietic stem cell：The fresh Cord blood of collection, passes through Ficoll lymphocyte separation mediums point
From mononuclearcell, after centrifugation, tunica albuginea layer is collected, plus physiological saline is resuspended and centrifuged again；Add program control after cell-protecting
Cooling, is stored in liquid nitrogen；
(4) before induction differentiation, umbilical cord blood hematopoietic stem cell is thawed, plus physiological saline is resuspended washing and centrifuged again, and collection is obtained
Umbilical cord blood hematopoietic stem cell；
(5) umbilical cord blood hematopoietic stem cell for obtaining step (4) is inoculated in 6 well culture plates by equal densities, and is added
The defined medium containing resveratrol that the step of equivalent (2) obtains；At 37 DEG C, gas concentration lwevel is under conditions of 5%
Culture；Carry out cell count respectively at the 1st~14 day and collect the cell progress flow cytomery of suspension growth.
The purity of resveratrol described in step (1) is >=99%, GC；Resveratrol stock solution concentration is
10mmol/L, the concentration is storage concentration, and working concentration is consistent with storage concentration, must be positioned over 4 DEG C of shading preservations.
Cell-protecting described in step (1) and (3) is preferably dimethyl sulfoxide (DMSO) (DMSO) and D-40
(Dextron) by volume 1:1 mixed liquor being mixed to get.
Defined medium described in step (2) includes Stemspan culture mediums, the IL-3 (20ng/ of STEMCELL companies
ML), IL-6 (50ng/mL), SCF (50ng/mL) and TPO (50ng/mL) cell factor, cell factor full name used is respectively
Thrombopoietin (Thrombopoietin, TPO), human interleukin -3 (Interleukin-3, IL-3), people are thin in vain
Born of the same parents' interleukin -6 (Interleukin-6, IL-6), human stem cell growth (Stem cell factor, SCF), totally 4 kinds of cells
The factor.
The concentration gradient of resveratrol is 5~25 μ in the defined medium containing resveratrol described in step (2)
mol/L；More preferably 15 μm ol/L；
Umbilical cord blood hematopoietic stem cell described in step (4), recovery of being thawed before differentiation culture is oriented from liquid nitrogen.
In umbilical cord blood hematopoietic stem cell into megakaryoblast directed differentiation incubation, add while adding cell factor
Enter RV, reach the effect of increase megakaryoblast directed differentiation quantity, of the invention is easy to operate, and cost is low, obtains
The megakaryoblast arrived is safe.
The preparation method of the present invention during Cord blood is external to megakaryoblast directed differentiation culture by adding
Resveratrol, the 7th day, the 14th day detection megakaryoblast growing amount.The present invention is carried by adding cheap resveratrol
The directed differentiation efficiency of high megakaryoblast.
The present invention has the following advantages and effect relative to prior art：
(1) present invention effectively increases umbilical cord blood hematopoietic stem cell to the quantity of megakaryoblast directed differentiation, finally obtains
A greater amount of megakaryoblasts are obtained, it is with low cost.
(2) defined medium containing resveratrol that the present invention is used is resveratrol culture medium, is exclusive configuration
Defined medium, being somebody's turn to do the defined medium containing resveratrol makes megakaryoblast directed differentiation operate more handy and safe and height
Effect.
(3) resveratrol stock solution prepared of the present invention, be resveratrol be dissolved in dimethyl sulfoxide (DMSO) (DMSO) and
D-40 (Dextron) volume ratio is 1:In 1 cell-protecting.Dimethyl sulfoxide (DMSO) is conventional organic solvent,
But there is certain toxic action to cell, D-40 has protective effect to cell, the white black false hellebore that this kind of method is prepared
Alcohol storing liquid is used for cell culture, not only ensure that the performance of resveratrol drug effect in cell cultivation process, and less
Toxic action of the dimethyl sulfoxide (DMSO) to cell.
Brief description of the drawings
Under Fig. 1 is the effect of each concentration resveratrol culture medium, each sample of umbilical cord blood hematopoietic stem cell is the 1st, 7,14 days
CD41+The result figure of surface antigen expression.
Under Fig. 2 is the effect of each concentration resveratrol culture medium, each sample of umbilical cord blood hematopoietic stem cell is the 1st, 7,14 days
The result figure of TCS amount.
Under Fig. 3 is the effect of each concentration resveratrol culture medium, each sample of umbilical cord blood hematopoietic stem cell is the 1st, 7,14 days
The result figure of cell quantity multiplied ratio.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited
In this.
Embodiment 1
Cord blood picks up from the pregnant baby of healthy puerpera.After testing hepatitis B, hepatitis C, syphilis, AIDS, cytomegalovirus,
TORCH detections, mycoplasma, Chlamydia, G-6PD and ground it is poor be feminine gender.Sample collects the traffic condition guarantor for transporting blood bank back certainly
Hold at 4~8 DEG C.Megakaryoblast directed differentiation operation is carried out by the following method：
1st, the preparation of resveratrol solution and collective media
Resveratrol (>=99%, GC), is dissolved in cell-protecting, is configured to concentration and is stored up for 10mmol/L resveratrols
Deposit solution.
Described cell-protecting is DMSO and Dextron by volume 1:1 mixed liquor being mixed to get.
Defined medium：The Stemspan culture mediums of STEMCELL companies, respectively addition IL-3 (20ng/mL), IL-6
(50ng/mL), SCF (50ng/mL) and TPO (50ng/mL) cell factor.
Defined medium containing resveratrol：The resveratrol stock solution of above-mentioned preparation is added to defined medium
In, obtain the defined medium containing resveratrol；Resveratrol final concentration of 0,5 μm of ol/L, 15 μm of ol/L, 25 μm of ol/
L；Wherein, final concentration of the 0 of resveratrol is control group (Control group).
2nd, the preparation of umbilical cord blood hematopoietic stem cell
Transport to obtain into fresh and healthy human cord blood of the storehouse in 4 hours from collection and be rich in CD34+Candidate stem cell
Mononuclearcell layer, Programmed freezing is frozen in liquid nitrogen and preserved；Thaw and recover before induction differentiation, be then inoculated in 6 orifice plates,
Then the defined medium containing resveratrol is added.Detailed process is as follows：
(1) fresh Cord blood is gathered, Guangdong Province's Cord Blood Bank is transported back in 4 hours, passes through Ficoll lymphocytes point
Chaotropic separates mononuclearcell, after centrifugation, collects tunica albuginea layer (tunica albuginea layer is in the middle of plasma layer and separation liquid layer), plus physiology
Washing is resuspended in salt solution, adds after cell-protecting, cools through Programmed freezing instrument and freezen protective is in liquid nitrogen.
(2) umbilical cord blood hematopoietic stem cell, which thaws, recovers, and is transferred to centrifuge tube, adds 3 times or so mL normal salines, washing
1 time, centrifugal force 200g is centrifuged 8 minutes, is collected cell precipitation, is produced umbilical cord blood hematopoietic stem cell；Umbilical cord blood hematopoietic stem cell
It is seeded on 6 well culture plates, adds the defined medium containing resveratrol so that the white black false hellebore of each sample correspondence respective concentration
Alcohol.
3rd, the identification of megakaryoblast
The identification of megakaryoblast is by the following method
(1) morphological analysis of megakaryoblast and cell count：The cell of culture is observed into cell in inverted microscope
State is simultaneously taken pictures.Few cells suspension is taken, cell quantity is calculated.
(2) phenotype test of flow cytometer：Take respectively after culture the 1st day, the 7th day, the cell of 14 days carried out CD41+
Detection.
Stem cell qualification result and cell activation assay：
1. the umbilical cord blood hematopoietic stem cell of the 1st day after observation is inoculated with inverted microscope, cell quantity is sufficient, and form is more
Single, in dispersity, cell volume is big and justifies, and cell space is bright.Cell counts, Control and 5 μm of ol/L, 15 μm of ol/
L, the hematopoietic stem/progenitor quantity of 25 μm of each samples of ol/L resveratrols are respectively：3.0×106Individual/mL, 2.86 × 106Individual/
mL、2.52×106Individual/mL, 2.68 × 106Individual/mL；As shown in Figure 2.
By flow cytomery, in CD41+In terms of cell proportion, Control and 5 μm of ol/L, 15 μm of ol/L, 25 μ
The numerical value of each sample of mol/L resveratrols is 18.61%；As shown in Figure 1.
2. the umbilical cord blood hematopoietic stem cell of the 7th day breaks up situation after observation inoculation under inverted microscope, compared with the 1st day,
Cell quantity is increased slightly, and form is broken up, and is largely the bright hematopoietic stem/progenitor of dispersed cell space, fraction hair
Estrangedization, clustering, nucleus become big, and after birth is unintelligible, and arrangement is in closely lumps.Cell counts, Control and
5 μm of ol/L, 15 μm of ol/L, hematopoietic stem/progenitor quantity of 25 μm of each samples of ol/L resveratrols are respectively：4.06×106Individual/
mL、4.06×106Individual/mL, 4.03 × 106Individual/mL, 4.01 × 106Individual/mL；As shown in Figure 2.In contrast to the 1st day, cell quantity
Ratio is respectively：135%th, 142%, 160%, 150%；As shown in Figure 3.
By flow cytomery, in CD41+In terms of cell proportion, Control and 5 μm of ol/L, 15 μm of ol/L, 25 μ
The numerical value of each sample of mol/L resveratrols is respectively：4.83%th, 3.5%, 4.26%, 3.54%；As shown in Figure 1.
After 7 days cultivate, the sample of resveratrol culture medium is added, the increased number of cell is slightly above Control, but
In terms of streaming result, Control is higher than all resveratrol samples.
3. the umbilical cord blood hematopoietic stem cell of the 14th day breaks up situation after observation inoculation under inverted microscope, compared with the 7th day,
Cell quantity is significantly increased, and obvious differentiation occurs in form, and≤50% is the bright hematopoietic stem/progenitor of dispersed cell space, is exceeded
Half for differentiation megakaryoblast, it will be apparent that cluster assemble, arrangement closely be in lumps, kytoplasm extremely enrich, it is intracellular containing
A large amount of particles.Cell counts, Control and 5 μm of ol/L, 15 μm of ol/L, the hematopoiesis of 25 μm of each samples of ol/L resveratrols
Ancestral cells quantity is respectively：10.32×106Individual/mL, 14.29 × 106Individual/mL, 14.45 × 106Individual/mL, 13.73 × 106
Individual/mL；As shown in Figure 2.It is respectively equivalent in the 1st day original cell quantity of various kinds sheet：344%th, 499%, 573%,
512%；As shown in Figure 3.
By flow cytomery, in CD41+In terms of cell proportion, Control and 5 μm of ol/L, 15 μm of ol/L, 25 μ
The numerical value of each sample of mol/L resveratrols is respectively：16.05%th, 14.36%, 16.63%, 24.62%；As shown in Figure 1.
After 14 days cultivate, the sample of resveratrol is added, cell proliferating number amount is apparently higher than Control, especially
15 μm of ol/L resveratrol samples, cell cultivation effect is the most obvious.
In terms of streaming result, Control is close with 15 μm of ol/L resveratrol samples, 25 μm of ol/L resveratrol samples
CD41+Numerical value is higher, and 5 μm of ol/L resveratrol samples are minimum.
In summary, according to the method for the present invention, in umbilical cord blood hematopoietic stem cell to megakaryoblast directed expansion process
In, in CD41+Cell proportion is not less than in the case of Control, and the culture medium containing 15 μm of ol/L resveratrols is significantly improved
The cell quantity of amplification, so that the growing amount of megakaryoblast is added, and also operating process is simple, safety.
Embodiment 2
Umbilical cord blood hematopoietic stem cell is carried out to megakaryoblast directed expansion with reference to the method for embodiment 1：
1st day, Control and 5 μm of ol/L, 15 μm of ol/L, the hematopoietic stem/progenitor of 25 μm of each samples of ol/L resveratrols
Starting quantity is respectively：4.26×106Individual/mL, 4.0 × 106Individual/mL, 3.54 × 106Individual/mL, 3.88 × 106Individual/mL.
6th day, Control and 5 μm of ol/L, 15 μm of ol/L, the hematopoietic stem/progenitor of 25 μm of each samples of ol/L resveratrols
Quantity is respectively：3.34×106Individual/mL, 3.73 × 106Individual/mL, 4.27 × 106Individual/mL, 4.46 × 106Individual/mL；Through excessively stream
Formula cell instrument is detected, in CD41+In terms of cell proportion, as a result it is respectively：6.35%th, 8.18%, 8.45%, 8.54%.
12nd day, Control and 5 μm of ol/L, 15 μm of ol/L, 25 μm of each samples of ol/L resveratrols Hematopoietic Stem/ancestral it is thin
Born of the same parents' quantity is respectively：15.57×106Individual/mL, 16.36 × 106Individual/mL, 17.64 × 106Individual/mL, 11.39 × 106Individual/mL,
In contrast to the 1st day cell quantity, each sample growth ratio was respectively：365.6%th, 409.2%, 498.44%, 293.7%；Through
Overflow-type cell instrument is detected, in CD41+In terms of cell proportion, as a result it is respectively：31.78%th, 33.38%, 32.97%,
31.81%.
Shown according to the streaming results contrast of the 12nd day, Control and 5 μm of ol/L, 15 μm of ol/L, 25 μm of white black false hellebores of ol/L
The CD41 of each sample of alcohol+Cell proportion is close；In terms of cell multiplied ratio, 15 μm of ol/L resveratrols samples apparently higher than
Control, with statistical significance.
Add the cell sample that resveratrol is co-cultured, CD41+Cell proportion and Control are in the case of, 15 μm of ol/
The cell proliferating number amount of L and 5 μm of ol/L resveratrol sample is apparently higher than Control, wherein 15 μm of ol/L effects are optimal, conclusion
It is same as Example 1.
Above-described embodiment is preferably embodiment, but embodiments of the present invention are not by above-described embodiment of the invention
Limitation, other any Spirit Essences without departing from the present invention and the change made under principle, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (9)
1. a kind of method for improving Cord blood megakaryoblast directed differentiation quantity, it is characterised in that：It is dry thin with umbilical cord blood hematopoietic
Born of the same parents are sample cell, and differentiation culture is oriented in the defined medium containing RV；
The concentration gradient of RV is 5~25 μm of ol/L in the described defined medium containing RV.
2. according to the method described in claim 1, it is characterised in that：White multitude in the described defined medium containing RV
The concentration of reed alcohol is 15 μm of ol/L.
3. the method according to any one of claim 1~2, it is characterised in that：The described specific training containing RV
Foster base be containing cell factor 20ng/mL IL-3,50ng/mL IL-6,50ng/mL SCF and 50ng/mL TPO culture medium.
4. method according to claim 3, it is characterised in that：Described cell factor is added to STEMCELL companies
In Stemspan culture mediums.
5. the method according to any one of claim 1~2 or described in 4, it is characterised in that including step in detail below：
(1) resveratrol is dissolved in cell-protecting, is configured to resveratrol stock solution；
(2) defined medium is prepared, the resveratrol stock solution that step (1) is prepared is added in defined medium, obtains
Defined medium containing resveratrol；
(3) separation of umbilical cord blood hematopoietic stem cell：The fresh Cord blood of collection, is separated single by Ficoll lymphocyte separation mediums
Individual nucleus, after centrifugation, collects tunica albuginea layer, plus physiological saline is resuspended and centrifuged again；Programmed freezing after cell-protecting is added,
It is stored in liquid nitrogen；
(4) before induction differentiation, umbilical cord blood hematopoietic stem cell thaws, plus physiological saline is resuspended washing and centrifuged again, and collection obtains umbilical cord
Blood candidate stem cell；
(5) umbilical cord blood hematopoietic stem cell for obtaining step (4) is inoculated in 6 well culture plates by equal densities, and adds equivalent
The step of (2) obtained defined medium containing resveratrol, cultivated.
6. method according to claim 5, it is characterised in that：The condition of culture described in step (5) is in 37 DEG C, dioxy
Change concentration of carbon be 5% under conditions of cultivate 1~14 day.
7. method according to claim 6, it is characterised in that：Cell count is carried out within the 1st~14 day respectively in culture and is received
The cell for collecting suspension growth carries out flow cytomery.
8. method according to claim 5, it is characterised in that：
Cell-protecting described in step (1) and (3) is dimethyl sulfoxide (DMSO) and D-40 by volume 1:1 mixing
Obtained mixed liquor.
9. method according to claim 5, it is characterised in that：
Resveratrol stock solution concentration described in step (1) is 10mmol/L.
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| CN105062969B (en) * | 2015-09-07 | 2018-08-28 | 广州市天河诺亚生物工程有限公司 | A method of improving Cord blood megakaryoblast Differentiation Induction in vitro efficiency |
| CN105754939A (en) * | 2016-04-15 | 2016-07-13 | 广州市天河诺亚生物工程有限公司 | Method for improving directional differentiation effects on erythroid progenitor cells in umbilical cord blood |
| CN106730015A (en) * | 2017-01-06 | 2017-05-31 | 朱凯林 | A kind of preparation for beauty and preparation method thereof |
| CN110982779B (en) * | 2019-12-25 | 2020-12-29 | 广州市天河诺亚生物工程有限公司 | Method for improving utilization rate of umbilical cord blood |
| CN111053788B (en) * | 2019-12-25 | 2021-02-26 | 广州市天河诺亚生物工程有限公司 | Composition for adjuvant therapy of chronic hemorrhage and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101517070A (en) * | 2006-05-31 | 2009-08-26 | Styx有限责任公司 | Methods of selecting stem cells and uses thereof |
| CN101864396A (en) * | 2010-05-17 | 2010-10-20 | 中国人民解放军军事医学科学院野战输血研究所 | Method for inducing megakaryoblast and megakaryocyte in vitro |
| CN103330720A (en) * | 2013-07-18 | 2013-10-02 | 广州市天河诺亚生物工程有限公司 | Mixing stem cell injection and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130287750A1 (en) * | 2009-10-14 | 2013-10-31 | Cellect Biotechnology Ltd. | Method of selecting stem cells and uses thereof |
-
2015
- 2015-05-12 CN CN201510239696.8A patent/CN104789530B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101517070A (en) * | 2006-05-31 | 2009-08-26 | Styx有限责任公司 | Methods of selecting stem cells and uses thereof |
| CN101864396A (en) * | 2010-05-17 | 2010-10-20 | 中国人民解放军军事医学科学院野战输血研究所 | Method for inducing megakaryoblast and megakaryocyte in vitro |
| CN103330720A (en) * | 2013-07-18 | 2013-10-02 | 广州市天河诺亚生物工程有限公司 | Mixing stem cell injection and preparation method thereof |
Non-Patent Citations (5)
| Title |
|---|
| Comparative analyses of megakaryocytes derived from cord blood and bone marrow;RIKA MIYAZAKI等;《British Journal of Haematology》;20001213;第108卷;第602-609页 * |
| Resveratrol ameliorates TNFa-mediated suppression of erythropoiesis in human CD34+ cells via modulation of NF-k signaling;Jee-Yeong Jeong等;《British Journal of Haematology》;20111231;第155卷;第93-101页 * |
| Resveratrol Increases the Bone Marrow Hematopoietic Stem and Progenitor Cell Capacity;Pauline Rimmelé等;《Am J Hematol》;20141231;第89卷(第12期);摘要 * |
| 脐血巨核祖细胞的体外扩增;冯义等;《中国医学科学院学报》;20050430;第27卷(第2期);第199-205页 * |
| 采用二甲基亚砜和右旋糖酐冻存脐血造血细胞的研究;刘斌等;《中华儿科杂志》;20001231;第38卷(第10期);第635-636页 * |
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