CN105820999A - Preparation for mobilizing mesenchymal stem cells - Google Patents

Preparation for mobilizing mesenchymal stem cells Download PDF

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CN105820999A
CN105820999A CN201610323312.5A CN201610323312A CN105820999A CN 105820999 A CN105820999 A CN 105820999A CN 201610323312 A CN201610323312 A CN 201610323312A CN 105820999 A CN105820999 A CN 105820999A
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preparation
dfo
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mscs
mesenchymal stem
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谢海涛
李相鲁
张严冬
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Guangdong Wanhai Cell Biotechnology Co Ltd
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
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Abstract

The invention provides a preparation for mobilizing mesenchymal stem cells .The preparation comprises DFO, and the dosage range of the DFO is 1-15 mg/kg .Preferably, the DFO and BOP are combined to be used .The preparation has the advantages that the mobilizing period is short, and the mesenchymal stem cells of a supplier can be efficiently collected within one hour or several hours; compared with a traditional method, the method has no side effect, and the supplier has no adverse effects such as skeleton pain and spleen swelling.

Description

A kind of preparation mobilizing mesenchymal stem cells MSCs
Technical field
Biological technical field of the present invention, is specifically related to a kind of preparation mobilizing mesenchymal stem cells MSCs.
Background technology
Mescenchymal stem cell (mesenchymalstemcells, MSCs) is the adult stem cell with powerful multiplication capacity and multi-lineage potential.Owing to the tissues such as bone, cartilage, fat, muscle, flesh is dirty, neural can be divided under certain condition, MSCs oneself become organizational project and the most potential seed cell of regenerative medicine.MSCs potential applicability in clinical practice is wide: treatment bone, joint, the dirty defect of flesh and damage；Promote neurocyte, myocardial cell, liver cell regeneration；Hematopoiesis support, promotes that hematopoietic stem cell is implanted；Graft versus host disease CGVHD after prevention and treatment hematopoietic stem cell transplantation)；Immune modulating treatment autoimmune disorder disease etc..
Bone marrow is the source that MSCs enriches the most, current clinical trial with in application, from donor or self to extract a certain amount of bone marrow, Isolation and culture be the main path obtaining MSCs.The feasible easy grasp of the method, but there is also some shortcomings: cell derived is limited, incubation step is loaded down with trivial details, amplification to required cell concentration requires time for longer, the non-humanization of culture fluid additive, pollution problem, tumorigenesis risk etc..The shortcomings of transplantation treatment after the foundation of mobilizing method can avoid In vitro culture in MSCs body, new way is opened up in the clinical practice for MSCs.
Stem cell is entered the process of external circulation by basic stitch site to mobilize (Mobilization) to refer to.Stem cell mobilization is the most potential methods for the treatment of diseases, and such as hematopoietic stem cell mobilization, oneself is widely used in clinic at present.Mescenchymal stem cell is mobilized and is referred to apply mobilization agent or use mobilization measure to promote MSCs in bone marrow discharge and move to peripheral blood.Effective MSCs mobilizes cell contamination that after can avoiding In vitro culture, infusion of therapeutic exists, incubation time is long easily misses the shortcomings such as window golden hour.Mobilize in MSCs body and transplant incomparable low wound and high efficiency after there is In vitro culture, therefore the most meaningful to the research of mobilizing method in MSCs body.Many researcheres are the most once attempted using granular leukocyte colony stimulating factor (G-CSF) to mobilize MSCs, but result is disappointing.The most still lack effectively clinical feasible MSCs mobilization agent.
In early-stage Study, it has been found that hypoxia-inducible factor-1 alpha (HIF-1 α) is the key factor (StemCellsDev.201120 (11): 1961-71) in mescenchymal stem cell mobilization.They establish the animal model that hypoxia inducible rat MSCs mobilizes, the kinetics mobilized by detection hypoxia inducible MSCs, it has been found that short-term anoxia can mobilize rat MSCs, through identifying the immunophenotype mobilizing entrance peripheral blood MSCs and skeletonization, one-tenth fat, becoming cartilage ability and derived from bone marrow MSCs basic simlarity.In further Mechanism Study, HIF-1 α plays pivotal role in hypoxia inducible MSCs mobilizes to have research worker to confirm, and HIF-1a downstream passages Stromal cell-derived factor-1α (SDF-1a) and receptor CXCR 4 thereof participate in MSCs and mobilize.HIF-1 is a heterodimer including HIF-1 α and two subunits of HIF-1 β.HIF-1 β is the subunit that many transcription factor are common, expresses in constitutive character intracellular.HIF-1a is then regulation subunit and the active subunits of HIF-1.The physiologically active of HIF-1 depends primarily on activity and the expression of HIF-1 α subunit.The regulation mainly level after protein translation that HIF-1 alpha expression amount increases, i.e. by increasing the stability of albumen, suppresses its degraded to realize.When cell is in normal oxygen condition, at prolyl hydroxylase (prolyl Hydroxylase, PHD) effect under, in HIF-1 alpha molecule, the proline residue of 402 and 564 is hydroxylated.Hydroxylated HIF-1a can be with tumor suppressor protein von Hippel Lindau (VHL) combines, and the latter can be identified generation ubiquitination degraded.In this degradation pathway, PHD is the enzyme played a crucial role.PHD belongs to Fe2+, the intoxicated 1,3-propanedicarboxylic acid of 2-rely on dioxygenase superfamily, be the rate-limiting enzyme of HIF-1a degradation reaction.PHD inhibitor can stablize the expression of HIF-1 α.Research finds, iron chelating agent Cobalt Chloride (CoCl2) the clean property of PHD can be suppressed, and suppress the hydroxylation of HIF-1 α, reduce the degraded of HIF-1 α under normal oxygen condition, be referred to as " anoxia simulant ".
SDF-1/CXCR4 axle plays an important role in mobilizing in stem cell body.Stroma cell derivative factor (SDF-l) is the CXC type chemotactic factor produced by marrow stromal cell, and CXC race cytokine receptor 4 (CXCR4) is unique receptor of SDF-1 known today.Research confirm, SDF-1 at endotheliocyte by the direct regulation and control of transcription factor HIF-1.AMD3100 is the small molecular antagonists of SDF-1 α/CXCR4 axle, it is possible to effectively block the interaction of SDF-1 α Yu CXCR4 and CXCR4 is not had agonism.
Although root understands HIF-1/SDF-1 α-CXCR4 axle with early-stage Study result according to the literature and plays a crucial role in MSCs mobilizes, but still lack a kind of effective MSCs at present and mobilize preparation, still lack feasible MSCs mobilizing method.
201310057178.5 1 kinds of preparations mobilizing mescenchymal stem cell of Chinese patent application and separation method thereof, have employed CoCl2Mobilizing mesenchymal stem cells MSCs with AMD3100, there is the feature of cycle length in this scheme, and CoCl2 Toxicity is relatively strong, damages patient body unavoidably.To this end, explore and study the key that reagent that is harmless and that mobilize the cycle short is research and development at present.
Summary of the invention
The purpose of the present invention is primarily directed to the problems of the prior art, now provides a kind of preparation mobilizing mesenchymal stem cells MSCs and separates and collects mescenchymal stem cell method.
For realizing the purpose of the present invention, the invention provides techniques below scheme:
A kind of preparation mobilizing mesenchymal stem cells MSCs, the preparation of described mobilization mesenchymal stem cells MSCs includes DFO.
The dosage range of described DFO is 1-15mg/kg.
Described DFO and BOP is used in combination.
Described DFO and AMD3100 is used in combination.
The dosage of described AMD3100 is 1-4mg/kg.
Described DFO and AMD3100 and BOP is used in combination.
The dosage of described BOP is 1-20mg/kg.
The dose ratio of described DFO, AMD3100 and BOP is: 1-3:1:1-4.
A kind of method utilizing the preparation mobilizing mesenchymal stem cells MSCs to separate and collect mescenchymal stem cell, the preparation described in utilization promotes mescenchymal stem cell to mobilize entrance peripheral blood then to separate.
The step separating and collecting mescenchymal stem cell is as follows: take peripheral blood, with lymphocyte separation medium separation mononuclearcell, resuspended by the DMEM culture medium containing 5-30% hyclone, it is seeded to culture bottle, changing liquid after cultivating 3-8 days, obtains peripheral blood mescenchymal stem cell for 9-15 days.
Described peripheral blood mescenchymal stem cell high expressed CD90, CD29, CD44, does not express CD45, CD34, has external skeletonization, becomes fat, one-tenth cartilage differentiation ability.
It is an advantage of the current invention that:
1, test proves that, the preparation mobilizing mesenchymal stem cells MSCs of the present invention mobilizes the cycle short, it is only necessary within 1 hour to a few hours, just can efficiently collect donor bone marrow stem cell；
2, compared with traditional method, the method has no side effect, and donor is without untoward reaction such as skeleton pain, spleen enlargements.
Detailed description of the invention
In order to illustrate in greater detail the present invention, be given and following prepare example.But the scope of the present invention is not limited thereto.
DFO powder normal saline is made into the solution that concentration is 10mg/ml, and AMD3100 normal saline is made into the solution of 1mg/ml, and BOP crystal normal saline concentration is the solution of 10mg/ml.Then using normal saline different solutions, the most all packets is all with rat as experimental subject,
DFO mobilization group: give rats by intraperitoneal injection DFO solution 10mg/kg every day；
DFO combines AMD3100 mobilization group: first give DFO 10mg/kg/d*7d, the 8th day lumbar injection AMD3100 5mg/kg；
DFO combines BOP mobilization group: every day gives DFO10mg/kg/d*7d and BOF10mg/kg/d*7d；
DFO combines BOP and AMD3100 mobilization group: first give DFO10mg/kg/d*7d and BOF10mg/kg/d*7d, the 8th day lumbar injection AMD3100 5mg/kg；
Gather each group of rat peripheral blood, use the quantity of two kinds of method each group of peripheral blood MSCs of detection of fibroblast-like cells Colony forming culture method (CFU-F method) and FCM analysis, it was demonstrated that DFO, DFO associating AMD3100 and DFO combines the BOP mobilized effects to MSCs.
Peripheral blood mobilization gone out becomes fiber-like colony to carry out Secondary Culture, by its surface markers CD44 of Flow cytometry, the expression of CD29, CD90, CD34, CD45, identifies immunophenotype.
Peripheral blood mobilization gone out becomes fiber-like colony to carry out Secondary Culture, carries out into fat, skeletonization, the differentiation of one-tenth chondrocyte induction, identifies the Multidirectional Differentiation ability of mobilized peripheral blood source MSCs.
Embodiment 1
1. preparation of reagents
DFO: weigh 100mgDFO powder, adds 10ml normal saline, is made into the storage liquid of 10mg/ml, subpackage after 0.22 μm filter filtration ,-20 DEG C of preservations.Concentration is adjusted to 4mg/ml when being administered by storage liquid normal saline dilution 10 by animal again.First the DFO solution of 3mg/ml, then preparation 2mg/ml DFO and 2mg/ml are prepared The mixed solution of BOP.
(2) AMD3100:5g powder adds 5ml normal saline, is made into the solution of 1mg/ml, and 0.22 μm filter filters subpackage ,-20 DEG C of preservations.Normal saline dilution 4 is used for mice to 1.25mg/ml again and is administered.
2. animal packet
(1) combining the AMD3100 mobilized effects to MSCs for detection DFO and DFO, rat is divided into 6 groups, often group 5, is respectively as follows: 1. normal saline group i.e. NS group: give rats by intraperitoneal injection normal saline (with DFO same volume) every day, totally 7 days；2. DFO7d group: give lumbar injection DFO solution 10mg/kg every day, totally 7 days；3. AMD3100 group: gives respective volume normal saline for 1-7 days, the 8th day lumbar injection AMD3100 5mg/kg gathers peripheral blood on time after 1 hour；4. DFO combines AMD3100 group: first give DFO 10mg/kg/d*7d, the 8th day lumbar injection AMD3100 5mg/kg；5. DFO combines BOP group: give DFO and BOP 5mg/kg/d respectively every day；6. DFO, BOP combine AMD3100 group: first give DFO and BOP 5mg/kg/d*7d, the 8th day lumbar injection AMD3100 5mg/kg respectively；Peripheral blood is gathered on time after 1 hour.
3. rat peripheral hemocyte extracts
After each process group arrives the respective handling time, take PERIPHERAL BLOOD MONONUCLEAR CELL respectively and carry out CFU-F method detection MSCs quantity and Flow cytometry CD45-CD90+Cell mass ratio.
4% chloral hydrate 10ml/kg intraperitoneal injection of anesthesia SD rat, takes blood 8-10ml, lymphocyte separation medium separation mononuclearcell (MNCs) with 10ml syringe from posterior vena cava, draws tunica albuginea layer, after PBS washs 3 times, and counting.
4.CFU-F method
MNCs 5ml contained in 6ml peripheral blood is contained 20%FBS, 1% mycillin LG-DMEM culture medium resuspended, being inoculated in floor space is 12.5cm2Plastic culture bottle in, be placed in 37 DEG C containing 5%C02Incubator is cultivated.Change liquid after cultivating 7 days, observe under inverted microscope, in the 10th day time, count peripheral blood CFU-Es.By in fibroblast sample form, grow intensive, > colony of 50 cells is designated as CFU-F.
The adherent i.e. amplification of one MSCs forms a CFU-F, theoretical based on this, CFU-F culture method oneself become the classical way of detection peripheral blood MSCs quantity.CFU-F method testing result finds: giving DFO10mg/kg/d*7d, rat peripheral blood CFU-Fs quantity relatively matched group dramatically increases.Compared with saline control group, in AMD3100 group peripheral blood, CFU-Fs quantity there is no substantially increase.Compared with alone DFO group, DFO associating AMD3100 can dramatically increase MSCs and mobilize efficiency.Applicant research finds: DFO10mg/kg gives can increase for 7 days the quantity of rat peripheral blood MSCs.DFO associating CXCR4 inhibitor AMD3100 can increase MSCs and mobilize efficiency.
5. CD45-CD90+ cell mass ratio in Flow cytometry rat peripheral blood and bone marrow
After counting, peripheral blood mononuclear cells of rats PBS adjusts cell concentration to 107/ mL adds cell after 100 μ L adjust and manages to 1.5mlEP, arranges monochromatic and double-colored Isotype control group simultaneously.Streaming antibody staining: CD90-PE often pipe 5 μ L, CD45-FITC often pipe 2 μ L.After lucifuge incubated at room 30min, PBS washs 2 times, with the resuspended rear addition streaming pipe of 400 μ LPBS, upper machine testing.
Flow cytometry finds: DFO10mg/kg gives 7 days to make CD45 in peripheral blood-CD90+Cell mass ratio raises, and sum increases, and sees following table.After DFO Yu AMD3100 combination, in rat peripheral blood, CD45-CD90+ cell mass ratio relatively DFO group is dialled further up, quantity the most substantially increases, after DFO Yu BOP combination, in rat peripheral blood, CD45-CD90+ cell mass ratio relatively DFO group is dialled further up, and quantity the most substantially increases, after DFO, BOP are combined with AMD3100, being dialled further up after CD45-CD90+ cell mass ratio relatively DFO associating BPO group and DOF with AMD3100 combination in rat peripheral blood, quantity the most substantially increases, and see table 1.This result also further demonstrate that DFO10mg/kg gives 7 days to mobilize with induced rat MSCs.DFO associating CXCR4 inhibitor AMD3100, BOP can increase MSCs and mobilize efficiency.Table 1 each process group peripheral blood mononuclear cells of rats quantity and CD45-CD90+Cell mass changes
6. flow cytometry peripheral blood and bone marrow CFU-Fs Immunophenotyping
Peripheral blood or derived from bone marrow CFU-Fs are expanded to P2 generation, 0.25% pancreatin EDTA digestion, and 1000r/min is centrifuged 6min, after PBS washes twice, and counting.Regulation concentration is to 5*105Cell/100 μ L, is divided into 7 pipes.2 pipe Control are separately added into FITC Yu PE Isotype control, and other 5 pipes are separately added into CD44-FITC2 μ L, CD29-PE2 μ L, CD90-PE5 μ L, CD45-FITC2 μ L, CD34-PE5 μ L.After temperature hatches 30min, PBS washs 2 times.It is placed in streaming pipe with 400 μ LPBS are resuspended, upper machine testing, Cell-Quest software analysis result.
The immunophenotype of Flow cytometry mobilized peripheral blood source MSCs (PB-MSCs), result display PB-MSCs does not express hemopoietic system labelling CD45 (0.20 scholar is O.10%), CD34 (0.37 scholar 0.38%), and positive expression CD44
(93.80 scholar 2.62%), CD29 (99.67 scholar 0.42%) and CD90 (98.50 scholar 2.18%), PB-MSCs expresses similar to derived from bone marrow MSCs surface antigen, identified, meets MSCs immunophenotype.
7. skeletonization moves towards differentiation qualification
Peripheral blood or derived from bone marrow CFU-Fs are expanded to P2 generation, 0.25% pancreatin EDTA digestion.By cell with 10,000/cm2It is inoculated in 6 orifice plates, often organizes 3 holes.Being placed in 37 DEG C to cultivate after 2 days containing 5%C02 incubator, change culture fluid, (LG-DMEM of 10%FBS is containing O.1 μM dexamethasone, 10mM β phosphoric acid Gan Bo, 50 μMs of vitamin to add Osteoblast Differentiation induction liquid.Within every 3 days, change liquid, cultivate 14 days, observation of cell form under inverted microscope.Osteoblast Differentiation result is identified in VonKossa dyeing: blot culture fluid in culture plate, 15min is fixed with 4% paraformaldehyde, distilled water rinses 2 times, and after naturally drying, every hole adds the 2% silver nitrate solution dyeing 5min that 2ml newly prepares, after deionized water fully washs, being placed in 15-30min under ultraviolet, distilled water rinses 2 times, and 5% sodium thiosulfate processes 2min, distilled water rinses 2 times, and Microscopic observation is taken pictures.Mobilized peripheral blood source MSCs (PB-MSCs) was induction differentiation the 14th day, and more calcium salt secretion i.e. occurs in cell surface, and VonKonssa dyeing is all positive, and these results suggest that PB-MSCs has Osteoblast Differentiation ability.
8. become fat to move towards differentiation to identify
Derived from peripheral blood CFU-Fs is expanded to P2 generation, 0.25% pancreatin EDTA digestion.By cell with 20,000/cm2It is inoculated in 6 orifice plates, often organizes 3 holes.It is placed in 37 DEG C containing 5%C02Incubator is cultivated, and when the nearly 90%-100% of cell merges, adds into fat induction liquid (containing 45mlLG-DMEM culture medium and 5ml Adipose Differentiation additive), within every 3 days, changes liquid, cultivate 21 days, observation of cell form under inverted microscope.Fat differentiated result is identified in oil red 0 dyeing: blotting culture fluid in culture plate, PBS washs 1-2 time gently, and 4% paraformaldehyde fixes 15min.After 60% isopropyl alcohol, every hole adds freshly prepared oil red 0 dye liquor 2ml, and dye 10min, blots after dye liquor again by 60% isopropyl alcohol 2 times, and deionization distilled water flushing 2 times, Microscopic observation is taken pictures.
Through induction differentiation 21 days, can be observed that mobilized peripheral blood source MSCs (PB-MSCs) endochylema all occurs that the high refractivity fat that cluster is distributed drips, dye in Chinese red through oil red 0.
9. become cartilage to move towards differentiation to identify
Peripheral blood or derived from bone marrow CFU-Fs are expanded to P2 generation, 0.25% pancreas enzyme-EDTA digestion.PBS washes twice, with cartilage differentiation liquid (serum-free HG-DMEM contains Sodium Pyruvate Insulin-Transferrin-selenium, O.1 μ L dexamethasone, 200 μ L vitamin Cs containing 1-).Re-suspended cell, 5*105/ pipe is inoculated in 15ml centrifuge tube, and often pipe liquor capacity is O.5ml, and 500g6min is centrifuged, and is eventually adding TGF-β 1(10ng/ml), TGF β 3(10ng/ml).It is placed in 37 DEG C containing 5%C02Incubator, changes liquid once in every 3 days, adds fresh cytokine every time.Cultivate the 21st day and observe the formation of chondrocyte agglomerate.
Toluidine blue staining identifies into cartilage differentiation result: cartilage differentiation cell mass fixes 24h with 4% paraformaldehyde, piece of tissue is taken out, wraps with silk stocking pouch, and silk thread fastens.Tissue dewatering, puts tissue agglomerate respectively in following solution: 50% ethanol 1h, 70% ethanol 15min, 80% ethanol 15min, 90% ethanol 15min and dehydrated alcohol 40min.Transparency of organization, puts agglomerate respectively in dimethylbenzene and ethanol geometric ratio mixed liquor 20min and pure dimethylbenzene 20min.Waxdip, embedding, then tissue slice (5mm/ sheet), tissue slice is dried overnight.Then dewax to water, put tissue agglomerate respectively in following solution: dimethylbenzene 30min, dimethylbenzene and ethanol geometric ratio mixed liquor 5min, dehydrated alcohol 10min, 95% ethanol 5min, 80% ethanol 5min, 70% ethanol 5min and 50% ethanol 5min, last deionization washing 5min.1% toluidine blue dye liquor dyes 10-15 minute, and flowing water rinses, and after conventional mounting, Microscopic observation is taken pictures.
We use external high density micelle culture method and Toluidine blue staining the originate mobilized peripheral blood one-tenth cartilage differentiation potential of MSCs (PB-MSCs) to be estimated.Found that: PB-MSCs breaks up 21 days through external one-tenth chondrocyte induction, and cell mass significantly increases before relatively breaking up.After cell micelle carries out embedding, section etc. processes, the expression of Toluidine blue staining detection chondrocyte characteristic secretory product acid sugar limb polysaccharide.Coloration result shows: after PB-MSCs differentiation, cell all colours in metachromasy, i.e. extracellular matrix is dyed to purple, and nucleus is navy blue.Result above shows: PB-MSCs has into cartilage differentiation potential.
Summary result, can be concluded that (1) can mobilize for DFO10mg/kg*7 days with induced rat MSCs: (2) DFO10mg/kg*7 days associating can increase MSCs for AMD31005mg/kg*7 time and mobilize efficiency: (3) mobilized peripheral blood source MSCs has the immunophenotype similar to bone marrow MSCs i.e. high expressed CD90, CD29, CD44, and do not express CD45, CD340 mobilized peripheral blood source MSCs have the Multidirectional Differentiation ability similar with bone marrow MSCs can to skeletonization, becomes fat, one-tenth chondrocyte induction to break up.
The above; being only the present invention preferably detailed description of the invention, but protection scope of the present invention is not limited thereto, any those of ordinary skill in the art are in the technical scope that the invention discloses; the change that can readily occur in or replacement, all should contain within protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with scope of the claims.

Claims (10)

1. the preparation mobilizing mesenchymal stem cells MSCs, it is characterised in that: the preparation of described mobilization mesenchymal stem cells MSCs includes DFO.
The preparation of mobilization mesenchymal stem cells MSCs the most according to claim 1, it is characterised in that: the dosage range of described DFO is 1-15mg/kg.
The preparation of mobilization mesenchymal stem cells MSCs the most according to claim 1, it is characterised in that: described DFO and BOP is used in combination.
The preparation of mobilization mesenchymal stem cells MSCs the most according to claim 1, it is characterised in that: described DFO and AMD3100 is used in combination.
The preparation of mobilization mesenchymal stem cells MSCs the most according to claim 4, it is characterised in that: the dosage of described AMD3100 is 1-4mg/kg.
The preparation of mobilization mesenchymal stem cells MSCs the most according to claim 4, it is characterised in that: described DFO and AMD3100 and BOP is used in combination.
7. according to the preparation mobilizing mesenchymal stem cells MSCs described in any one of claim 3 or 6, it is characterised in that: the dosage of described BOP is 1-20mg/kg.
The preparation of mobilization mesenchymal stem cells MSCs the most according to claim 7, it is characterised in that: the dose ratio of described DFO, AMD3100 and BOP is: 1-3:1:1-4.
9. the method that the preparation utilized described in any one of claim 1-8 separated and collected mescenchymal stem cell, it is characterised in that: the preparation described in utilization promotes mescenchymal stem cell to mobilize entrance peripheral blood then to separate.
Separation the most according to claim 9 the method collecting mescenchymal stem cell, it is characterized in that: the step separating and collecting mescenchymal stem cell is as follows: take peripheral blood, with lymphocyte separation medium separation mononuclearcell, resuspended by the DMEM culture medium containing 5-30% hyclone, it is seeded to culture bottle, changing liquid after cultivating 3-8 days, obtains peripheral blood mescenchymal stem cell for 9-15 days.
CN201610323312.5A 2016-05-17 2016-05-17 Preparation for mobilizing mesenchymal stem cells Pending CN105820999A (en)

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Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BENJAMIN CAO, ET AL: "Therapeutic targeting and rapid mobilization of endosteal HSC using a small molecule integrin antagonist", 《NATURE COMMUNICATIONS》 *
SANJAY KUMAR，ET AL: "Mobilization of bone marrow mesenchymal stem cells in vivo augments bone healing in a mouse model of segmental bone defect", 《BONE》 *
刘丽珍: "缺氧与缺氧模拟剂对间充质干细胞的动员作用及机理研究", 《中国博士学位论文全文数据库》 *
陈海旭等: "骨髓干细胞动员在心脑血管疾病中的研究进展", 《中国科学》 *

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Application publication date: 20160803