WO2012167588A1 - Séquence d'acides aminés d'un peptide ayant une affinité pour les cellules souches mésenchymateuses de la moelle osseuse, procédés de criblage et utilisation de ce peptide - Google Patents

Séquence d'acides aminés d'un peptide ayant une affinité pour les cellules souches mésenchymateuses de la moelle osseuse, procédés de criblage et utilisation de ce peptide Download PDF

Info

Publication number
WO2012167588A1
WO2012167588A1 PCT/CN2011/084339 CN2011084339W WO2012167588A1 WO 2012167588 A1 WO2012167588 A1 WO 2012167588A1 CN 2011084339 W CN2011084339 W CN 2011084339W WO 2012167588 A1 WO2012167588 A1 WO 2012167588A1
Authority
WO
WIPO (PCT)
Prior art keywords
bone marrow
mesenchymal stem
marrow mesenchymal
phage
stem cells
Prior art date
Application number
PCT/CN2011/084339
Other languages
English (en)
Chinese (zh)
Inventor
敖英芳
邵振兴
皮彦斌
张辛
周春燕
Original Assignee
北京大学第三医院
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 北京大学第三医院 filed Critical 北京大学第三医院
Publication of WO2012167588A1 publication Critical patent/WO2012167588A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus

Definitions

  • the invention belongs to the field of biomedicine, and particularly relates to an amino acid sequence, screening method and application of a bone marrow mesenchymal stem cell affinity polypeptide. Background technology
  • Tissue Engineering has been widely used in various clinical fields, including bone tissue, cartilage, nerves, blood vessels, skin, and regeneration and repair of the gastrointestinal and genitourinary systems.
  • tissue engineering In the development of tissue engineering, continuous improvement and renewal of the various organizations' supports is key, including material renewal, improvement of production methods and continuous innovation of construction concepts.
  • Another recent obvious change in the concept of tissue engineering is to "make it simple", that is, to simulate the composition of normal tissues as much as possible by various complicated means and techniques in vitro, and then implant In the body, it is better to provide a self-repairing stent for the human body, so that the natural "bioreactor" of the human body is attached to the stent for self-repair.
  • the human body environment is very complicated, it will lead to a lot of experimental ideas and results, and the complexity of the stent implantation step will limit its clinical promotion and application.
  • such scaffolds require specific modifications as needed to induce adhesion, proliferation and differentiation of specific cells.
  • the matrix surrounding the cells plays an important role in the biological function of the cells.
  • the functional domains of the proteins or polypeptide molecules in the matrix bind to the cell surface receptors, activate the complex signaling pathways in the cells, and express the genes of the cells.
  • Biological functions such as adhesion, migration, proliferation, and differentiation are regulated, and these domains may be only a few amino acid fragments in length.
  • This regulation method provides us with a theoretical basis for constructing active polypeptide sequences and surface modification of tissue engineering scaffolds to simulate the regulation function of the surrounding cells on the cells themselves. Modification of the scaffold with different types of polypeptides also confers different biological functions to the scaffold.
  • the modification of the scaffold by the cell-specific affinity polypeptide sequence can increase the adhesion rate of the corneal epithelial cells to the scaffold, and induce the stratification of the corneal cells on the scaffold, and the tissue structure similar to the physiological condition appears.
  • the modification of the scaffold by the polypeptide can also regulate the aggregation of the protein molecule on the tissue engineering scaffold.
  • Ryadnov et al. use a small molecular protein constructed by a plurality of polypeptide fragments to modify the scaffold, and use the polypeptide to exhibit a pro-specific molecular molecule.
  • the stent When the degree of modification is 5% -10% At the 4th week after implantation, the stent still exerts an excellent sustained release function of growth factors. And most importantly, there was no significant difference in the final repair of cartilage defects between the explants of exogenous TGF- ⁇ and the scaffolds that were not transfected with exogenous TGF- ⁇ , suggesting that the TGF- ⁇ affinity peptide modified scaffold, In the absence of exogenous growth factors, endogenous TGF- ⁇ growth factor can be enriched in a large amount, and the microenvironment in the scaffold can be improved to achieve the function of inducing differentiation of stem cells into cartilage.
  • Bone marrow mesenchymal stem cells (BMSCs), a kind of stem cells with multi-directional differentiation potential in bone marrow tissue, were proposed by Friedenstein et al in the 1960s, and their cytological properties have been studied. A large number of related research results have been published, especially in the past two decades. With the rapid development of tissue engineering and regenerative medicine, the demand for seed cells has led to a deeper research on various stem cells. At present, the multi-directional differentiation potential of bone marrow mesenchymal stem cells has been confirmed by a large number of studies, and because of its strong proliferative activity and multi-directional differentiation characteristics, it has been widely used in myocardium, nerve, liver, pancreas, Bone tissue and cartilage regeneration in all areas of tissue engineering.
  • the object of the present invention is to provide a screening method for a bone marrow mesenchymal stem cell affinity polypeptide capable of improving the specific affinity of a biological material for bone marrow mesenchymal stem cells in view of the above-mentioned drawbacks of the prior art.
  • Another object of the invention is to provide an amino acid sequence of a bone marrow mesenchymal stem cell affinity polypeptide.
  • a further object of the invention is to provide the use of a bone marrow mesenchymal stem cell affinity polypeptide.
  • a screening using improved phage display technology can enhance the specificity of biological materials for bone marrow mesenchymal stem cells
  • a method for the ability of a bone marrow mesenchymal stem cell affinity polypeptide comprising the following steps:
  • Primary culture of human bone marrow mesenchymal stem cells and human fibroblasts Primary cultured cells were obtained by primary culture of human bone marrow mesenchymal stem cells and passaged one generation; primary cultured and propagated by human fibroblasts Above generation, obtain negative screening cells;
  • Negative screening a phage library is added to the negative screening cells to remove the polypeptide fragment bound to the P10 generation ACL fibroblasts in the phage library;
  • Phage amplification The bone marrow mesenchymal stem cells obtained in the step 3 are extracted, and a cell lysate is prepared, and the phage titer in the cell is amplified;
  • Another technical solution of the present invention is an application of a bone marrow mesenchymal stem cell affinity polypeptide modified to a human scaffold and a modification method thereof, comprising the following steps: 3' C-end of the selected bone marrow mesenchymal stem cell affinity polypeptide , a cysteine (C) is attached, and the cysteine (C) residue is covalently coupled with the surface of the polyhexanolide (PCL) electrospinning membrane by the ammoniated surface -NH 2 to make PCL
  • the nanofiber membrane scaffold has the property of specifically enriching bone marrow mesenchymal stem cells.
  • PCL nanofiber membrane was placed in 10% w/v 1,6-hexanediamine in an isopropyl alcohol configuration at 37 ° C for 1 h;
  • Bone marrow mesenchymal stem cell-specific affinity polypeptide fragments are obtained by phage display technology, so that they can be highly targeted to bone marrow mesenchymal stem cells.
  • the surface of the PCL nanofiber membrane is modified by covalently coupling the affinity polypeptide fragment with the PCL electrospinning membrane to specifically enrich the bone marrow mesenchymal stem cells, so that the material is used as a scaffold
  • the bone marrow mesenchymal stem cells can be continuously adhered and a normal extracellular matrix (ECM) is produced, thereby achieving a better repair effect.
  • ECM extracellular matrix
  • the polypeptide has no obvious species specificity, so the affinity polypeptide sequence can be widely used in various cytological experiments and animal experiments as a
  • the affinity vector for screening bone marrow mesenchymal stem cells can screen and purify bone marrow mesenchymal stem cells more efficiently.
  • Example 1 is a diagram showing the results of primary culture of human bone marrow mesenchymal stem cells provided in Example 1 of the present invention
  • 2a, 2b, and 2c are the results of the phage blue spot assay provided in the phage titer assay of Example 1 of the present invention; 10 ⁇ l of the phage diluted 10 ⁇ 1 was added to 200 ⁇ l of Escherichia coli to incubate 1 After 5 minutes, the top medium was added and rapidly mixed, and the tetracycline-resistant LB-tet plate was plated at 37 ° C, 5% CO 2 overnight to calculate the number of phage blue spots on the plate. This number is then multiplied by the dilution factor to obtain the plaque forming unit (pfu) titer per 10 ⁇ phage. A single blue spot was picked up and amplified in LB/E. coli culture medium, and phage DNA sequencing was performed.
  • Figure 2a The number of blue spots is 10°
  • Figure 2b the number of blue spots is 10 1
  • Figure 2c The number of blue spots is 10 2 .
  • Figure 3 is a graph showing the recovery rate of four rounds of screened phage provided in the screening of phage affinity polypeptides of Example 1 of the present invention
  • Figure 4a is a graph showing the results of flow cytometry of human MSC affinity polypeptides provided in Example 2 of the present invention
  • Figure 4b is a graph showing the results of quantitative analysis of the fluorescence intensity of Figure 4a;
  • Figure 5a is a graph showing the results of flow cytometry of a rat MSC affinity polypeptide provided in Example 2 of the present invention
  • Figure 5b is a graph showing the results of quantitative analysis of the fluorescence intensity of Figure 5a;
  • Figure 5c is a graph showing the results of flow cytometry of the MSC affinity polypeptide of rabbits provided in Example 2 of the present invention
  • Figure 5d is a graph showing the results of quantitative analysis of the fluorescence intensity of Figure 5c;
  • rat (Rat) FITC-labeled mismatched polypeptide was incubated with rat bone marrow mesenchymal stem cells with an average fluorescence intensity of 5; FITC-labeled affinity polypeptide and rat bone marrow mesenchymal stem cells Co-incubation with an average fluorescence intensity of 65.
  • the fluorescence intensity of the affinity peptide group (65) is much higher than that of the mismatched peptide group (5);
  • the left peak represents the mismatched polypeptide and the right peak represents the affinity polypeptide.
  • Counts is the quantity.
  • the average fluorescence intensity in Figure 4b, Figure 5b, and Figure 5d is the average fluorescence intensity.
  • Figure 6 is a result of laser confocal microscopy observation provided by Example 2 of the present invention; the affinity polypeptide and mismatch polypeptide labeled with FITC are separately incubated with bone marrow mesenchymal stem cells for lh, phalloidin counterstained cytoskeleton, harness counterstaining The nucleus and laser confocal microscopy were used to observe the binding of cells to peptides.
  • the results showed that the affinity polypeptide sequence (EPLQLKM) had significantly higher affinity for bone marrow mesenchymal stem cells than the mismatched polypeptide (MLKPLEQ);
  • Figure 7 is a schematic view showing the attachment of an affinity polypeptide fragment to the surface of a polycaprolactone (PCL) nanoelectrospun fiber membrane by covalent bonding according to Example 3 of the present invention
  • Example 8 is a polypeptide linkage observed by a confocal microscope under the laser confocal microscope of a bone marrow mesenchymal stem cell-specific polypeptide modified PCL nanoelectrospun fiber membrane provided by Example 3 of the present invention
  • PCL Polycaprolactone
  • EPLQLKM FITC-labeled affinity peptide fragment
  • FIGS. 9a and 9b are the bone marrow mesenchymal stem cell-specific polypeptide-modified PCL nano-electrospins provided in Example 3 of the present invention.
  • the effect of the silk scaffold is a fragment of the affinity peptide of the bone marrow mesenchymal stem cell;
  • Figure 9b is a fragment of the mismatched polypeptide;
  • PCL Polycaprolactone
  • EPLQLKM bone marrow mesenchymal stem cell affinity peptide fragment
  • MLKPLEQ mismatched polypeptide fragment
  • Human bone marrow tissue was obtained from patients undergoing total knee arthroplasty (TKA), placed in a blood collection tube containing K2 EDTA anticoagulation (BD company), and washed with PBS (0.01 M, pH 7. 4). 2 times, the bone marrow tissue was evenly spread on the bottom of the culture dish, and added to DMEM (LG) complete medium containing 10% FBS, observed for 24 to 48 hours. After seeing the adherent cells, change the medium and continue the culture, then 2 ⁇ 3 Change the liquid once a day and pass it on for generations. As a positive screening cell (see Figure 1). Among them, DMEM is a medium containing various amino acids and glucose.
  • Human anterior cruciate ligament tissue was harvested from patients undergoing total knee arthroplasty (TKA), PBS (0.01 M, pH 7.4), membranous tissue and blood clots were removed, and cut with ophthalmic scissors. Broken, trypsin for 30 min at 37 °C to remove impurities, add complete medium to stop digestion, wash twice with PBS (0.01 M, pH 7.4), then add 0.2% of type I collagenase (formulated in DMEM) Digested at 37 ° C for 2-3 hours, shaken at 37 ° C per hour for 5 min, until most of the tissue blocks were visible to the naked eye. Under the inverted microscope, most of the cells were separated, and the elbow was light. Lightly blow, add an equal volume of complete medium to neutralize collagenase, centrifuge the digested cell suspension, discard the supernatant, resuspend in complete medium, plate, and pass for more than ten generations. As a negative screening cell.
  • PI generation source SC medullary mesenchymal stem cells
  • trypsinize wash PBS (0. 01M, pH 7. 4) 2 ⁇ 3 times; add blocking buffer to resuspend in lml EP tube
  • the cell was counted (2 X 10 6 ); the cell was counted (2 ⁇ 10 6 );
  • step (2) to obtain the negatively screened phage polypeptide library bacterial solution, and leave it at room temperature for 30 min;
  • Phage amplification see Ph. D. -7TM Phage Display Peptide Library Kit: a. Prepare the cell lysate from the positively screened cells obtained in step (3), and expand the phage titer in the cell. 5 ⁇ ; The cell lysate was added to a solution of the ER2738 (in the early-log), shaking at 37 ° C, incubated for 4.5 hours;
  • step 1) The culture solution in step 1) is added to a centrifuge tube, centrifuged at 10, OOO rpm for 10 minutes at 4 ° C, the supernatant is transferred to a new tube, and centrifuged again (10, OOO rpm, lOmin);
  • Phage titer determination (see Ph. D. -7TM Phage Display Peptide Library Kit): a. Inoculate ER2738 single colony in 5_10 ml LB medium, incubate at 37 ° C, shaker at 250 rpm to mid-log phase (0D 6 . . . ⁇ 0. 5).
  • the upper layer of agar is heated and melted in a microwave oven and divided into 3 ml/part into a sterile test tube, one tube of each dilution of the phage. Store at 45 ° C for later use.
  • the phage was diluted 10 fold in LB medium. Recommended dilution range: Amplified phage culture supernatant: 10 8 -10 u ; Unamplified panning eluate: ⁇ For each dilution, replace with a fresh tip. It is recommended to use a filter tip. Avoid cross contamination.
  • the phage-infected E. coli broth was added to a 45 °C pre-warmed top agar culture tube, mixed one time at a time, and immediately poured onto a pre-warmed LB/IPTG/Xgal plate at 37 °C. Properly tilt the plate to spread the upper agar evenly.
  • Plaque amplification Escherichia coli amplified bacterial solution (0D: 0.5) was diluted 1:100 with LB medium, and each plaque clone to be sequenced was dispensed into a tube, 1 ml/tube. ;
  • phage bacterial solution was transferred to a centrifuge tube and centrifuged for 30 seconds. The supernatant is transferred to a new tube and centrifuged. Pipette 80% of the supernatant into a new centrifuge tube. This is a solution for amplifying the phage. It can be stored at 4 °C for several weeks without much effect on the titer. Long-term storage application of sterile glycerol 1: 1 dilution, storage at -20 ° C; e. From the above amplified monoclonal phage liquid, absorb 500 ⁇ into a new centrifuge tube;
  • the pellet is resuspended in 20 ⁇ double distilled water, which is a phage template DNA solution;
  • Phage affinity peptide screening Using a phage display library (Ph.D. -7TM Phage Display Peptide Library Kit; New England Biolabs), a total of 4 rounds of screening were performed. After each screening, the phage titer was obtained and multiplied by The total volume of the bacterial liquid is the total amount of phage recovered after screening, and the selected phage is amplified, and the amplified phage is used for the next round of screening.
  • Phage affinity peptide screening Using a phage display library (Ph.D. -7TM Phage Display Peptide Library Kit; New England Biolabs), a total of 4 rounds of screening were performed. After each screening, the phage titer was obtained and multiplied by The total volume of the bacterial liquid is the total amount of phage recovered after screening, and the selected phage is amplified, and the amplified phage is used for the next round of screening.
  • the first round of screening Add phage stock solution (titer: 1 X10 13 PFU/10 1), after lysing the cells, extract the phage titer to 1X10 3 PFU /10 ⁇ 1, and after the extracted phage is amplified, the phage titer is 1X10 U PFU/I0 1;
  • the second round of screening Adding the amplified phage 10 ⁇ 1 , after screening, the recovered phage titer is 4.1 ⁇ 10 4 PFU /10 ⁇ 1, and the titer after amplification is 1 ⁇ 10 U PFU/I0 1
  • the third round of screening After screening, the recovered phage titer is 5.6 ⁇ 10 6 PFU /10 ⁇ 1, and the titer after amplification is 1 X 10 U PFU/10 ⁇ 1;
  • the fourth round of screening After screening, the phage titer is recovered. It is 3.0X10 5 PFU /10 ⁇ 1.
  • EPLQLKM bone marrow mesenchymal stem cell affinity phage
  • the phage recovery rate of the affinity-containing polypeptide 5.60E-06; the recovery rate of the phage stock solution: 1.00E-09
  • the obtained bone marrow mesenchymal stem cell affinity phage increased the affinity for bone marrow mesenchymal stem cells by 5600 times (5.60E-06/1.00E_09).
  • EPLQLKM EPLQLKM
  • the polypeptide sequence of 3 clones is: EPLQLKM; of the 20 phage clones determined in the third round, the polypeptide sequence of 7 clones is : EPLQLKM; of the 20 phage clones determined in the fourth round, the polypeptide sequence of 10 clones was: EPLQLKM.
  • the first round of screening results lacked specificity, so the polypeptide fragments carried by the phage in the eluate were not detected. Therefore, after four rounds of screening, a polypeptide sequence with high affinity for bone marrow mesenchymal stem cells was obtained: EPLQLKM.
  • Method Take a BMSC, and digest it with 0.05% EDTA for 0 minutes until the cells are completely detached. Collect the suspension, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and again The cells were suspended in PBS (0.01 M, pH 7.4), repeated 3 times, and the cells were filtered through a 400 mesh filter, and then centrifuged again (2000 rpm, 5 min), 0. lml PBS (0.01 M, pH 7. 4) was resuspended.
  • InM's SMSC affinity polypeptide (FITC-EPLQLKMC) and mismatched polypeptide (FITC-MLKPLEQC) were added, incubated for 1 h at room temperature, centrifuged (2000 rpm, 5 min), and washed 3 times with PBS (0.1 M, pH 7.4). 0. 5ml PBS (0. 01M, pH 7. 4) The cells were resuspended and transferred into a flow tube for flow cytometry analysis.
  • FITC-EPLQLKMC InM FITC-labeled bone marrow mesenchymal stem cell affinity peptide
  • FITC-EPLQLKMC InM FITC-labeled bone marrow mesenchymal stem cell affinity peptide
  • FITC mismatched polypeptide
  • the mismatched polypeptide was used as a blank control group with an average fluorescence intensity of 6; the primary bone marrow mesenchymal stem cells incubated with the bone marrow mesenchymal stem cell affinity polypeptide had an average fluorescence intensity of 189.
  • the fluorescence intensity of the bone marrow mesenchymal stem cell affinity polypeptide group was 31.5 times that of the mismatched polypeptide, indicating that the bone marrow mesenchymal stem cell affinity polypeptide has significant affinity for human-derived mesenchymal stem cell cells (see Figure 4a). And Figure 4b).
  • RESULTS Cell slides were prepared from human-derived mesenchymal stem cells, fixed in 4% paraformaldehyde for 10 min, washed three times with PBS, and FITC-affinity affinity peptides and mismatched peptides were incubated with cell slides for 1 h at 37 °C.
  • Bone marrow mesenchymal stem cell affinity polypeptides can accumulate in large amounts around and inside bone marrow mesenchymal stem cells, while only a very small number of mismatched random polypeptides are randomly endocytosed by bone marrow mesenchymal stem cells. This indicates that the bone marrow mesenchymal stem cell affinity polypeptide has a high affinity for human-derived mesenchymal stem cells (see Figure 6).
  • crosslinker 4- (N-maleimidomethyl)cyclohexane-1-carboxylic acid-3-sulfosuccinimide ester per well (1 mg/ml SMCC, Thermo Company), placed at room temperature for lh;
  • the affinity peptide fragment EPLQLKM and the mismatched polypeptide fragment MLKPLEQ were ligated to the surface of the polycaprolactone (PCL) nano-electrospinning scaffold by covalent binding (see Figure 7) and placed in a bone marrow mesenchymal stem cell suspension. Co-cultivation.
  • PCL nanofiber membrane to which the affinity polypeptide was linked was more favorable for the adhesion and growth of bone marrow mesenchymal stem cells than the PCL nanofiber membrane to which the mismatched polypeptide was ligated.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Biophysics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Transplantation (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Veterinary Medicine (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Dermatology (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne un peptide à heptades criblé par exposition sur phage, dont la séquence d'acides aminés est représentée par SEQ ID NO° 1, ledit polypeptide ayant une affinité spécifique pour les cellules souches mésenchymateuses de la moelle osseuse. La présente invention concerne également des procédés de criblage dudit peptide à affinité et son utilisation, notamment pour améliorer l'affinité d'une substance biologique pour les cellules souches mésenchymateuses de la moelle osseuse, et pour la réparation du cartilage en génie tissulaire.
PCT/CN2011/084339 2011-06-09 2011-12-21 Séquence d'acides aminés d'un peptide ayant une affinité pour les cellules souches mésenchymateuses de la moelle osseuse, procédés de criblage et utilisation de ce peptide WO2012167588A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201110153614.X 2011-06-09
CN 201110153614 CN102229646B (zh) 2011-06-09 2011-06-09 骨髓间充质亲干细胞亲和多肽的氨基酸序列、筛选方法及应用

Publications (1)

Publication Number Publication Date
WO2012167588A1 true WO2012167588A1 (fr) 2012-12-13

Family

ID=44842260

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2011/084339 WO2012167588A1 (fr) 2011-06-09 2011-12-21 Séquence d'acides aminés d'un peptide ayant une affinité pour les cellules souches mésenchymateuses de la moelle osseuse, procédés de criblage et utilisation de ce peptide

Country Status (2)

Country Link
CN (1) CN102229646B (fr)
WO (1) WO2012167588A1 (fr)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102229646B (zh) * 2011-06-09 2013-07-24 北京大学第三医院 骨髓间充质亲干细胞亲和多肽的氨基酸序列、筛选方法及应用
CN103877616B (zh) * 2014-03-18 2016-01-20 北京大学第三医院 一种软骨组织工程修复支架及其制备方法
CN106880871B (zh) * 2017-01-18 2019-12-13 烟台正海生物科技股份有限公司 一种用于促进子宫内膜修复的胶原蛋白真皮材料及其制备方法
CN107007883B (zh) * 2017-02-16 2020-02-07 北京大学第三医院 一种软骨修复支架及其制备方法
CN107551314A (zh) * 2017-09-21 2018-01-09 浙江大学 一种促进骨髓间充质干细胞粘附的e7‑胶原膜及其制备方法
CN109364295B (zh) * 2018-09-30 2021-08-24 上海交通大学医学院附属第九人民医院 丝素蛋白-多巴胺-e7短肽复合支架及其制备方法和应用
CN116271241A (zh) * 2021-12-09 2023-06-23 北京博辉瑞进生物科技有限公司 用于组织修复的改性非对称sis膜、其制备方法及应用
CN116899018B (zh) * 2023-09-13 2023-12-12 四川大学 E7多肽-明胶纳米纤维微球支架及其制备方法和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1978463A (zh) * 2005-12-09 2007-06-13 中国人民解放军军事医学科学院野战输血研究所 间充质干细胞亲和肽的筛选与用途
WO2009026218A2 (fr) * 2007-08-17 2009-02-26 University Of Illinois Identification et utilisation d'inhibiteurs peptidiques de la synthèse protéique
CN102229646A (zh) * 2011-06-09 2011-11-02 北京大学第三医院 骨髓间充质亲干细胞亲和多肽的氨基酸序列、筛选方法及应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101890184B (zh) * 2009-05-21 2013-07-10 四川大学华西医院 一种软骨源性胶原海绵支架及其制备方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1978463A (zh) * 2005-12-09 2007-06-13 中国人民解放军军事医学科学院野战输血研究所 间充质干细胞亲和肽的筛选与用途
WO2009026218A2 (fr) * 2007-08-17 2009-02-26 University Of Illinois Identification et utilisation d'inhibiteurs peptidiques de la synthèse protéique
CN102229646A (zh) * 2011-06-09 2011-11-02 北京大学第三医院 骨髓间充质亲干细胞亲和多肽的氨基酸序列、筛选方法及应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
XUE Z. X. ET AL.: "Preparation of electrospun fibrous vascular scaffolds with small diameter and the study of its blood compatibility.", JOURNAL OF FUNCTIONAL MATERIALS., vol. 40, no. 10, 20 October 2009 (2009-10-20), pages 1716 - 1719 *

Also Published As

Publication number Publication date
CN102229646B (zh) 2013-07-24
CN102229646A (zh) 2011-11-02

Similar Documents

Publication Publication Date Title
WO2012167588A1 (fr) Séquence d'acides aminés d'un peptide ayant une affinité pour les cellules souches mésenchymateuses de la moelle osseuse, procédés de criblage et utilisation de ce peptide
Qiu et al. Periosteal matrix-derived hydrogel promotes bone repair through an early immune regulation coupled with enhanced angio-and osteogenesis
Yan et al. Enhanced osteogenesis of bone marrow‐derived mesenchymal stem cells by a functionalized silk fibroin hydrogel for bone defect repair
Yang et al. Tooth root regeneration using dental follicle cell sheets in combination with a dentin matrix-based scaffold
Peng et al. Decellularized Disc Hydrogels for hBMSCs tissue-specific differentiation and tissue regeneration
Lesman et al. Engineering vessel-like networks within multicellular fibrin-based constructs
Peng et al. Genipin‐crosslinked decellularized annulus fibrosus hydrogels induces tissue‐specific differentiation of bone mesenchymal stem cells and intervertebral disc regeneration
Qiu et al. Cyclic tension promotes fibroblastic differentiation of human MSCs cultured on collagen‐fibre scaffolds
US9144585B2 (en) Isolated mesenchymal progenitor cells and extracellular matrix produced thereby
Sinthuvanich et al. Iterative design of peptide-based hydrogels and the effect of network electrostatics on primary chondrocyte behavior
Wang et al. Self‐assembled peptide‐based hydrogels as scaffolds for proliferation and multi‐differentiation of mesenchymal stem cells
Li et al. Coculture of peripheral blood CD34+ cell and mesenchymal stem cell sheets increase the formation of bone in calvarial critical-size defects in rabbits
Meng et al. Microfracture combined with functional pig peritoneum-derived acellular matrix for cartilage repair in rabbit models
KR20120092558A (ko) 시험관내 및 생체내 연골형성을 위한 방법 및 조성물
Shao et al. Surface modification on polycaprolactone electrospun mesh and human decalcified bone scaffold with synovium‐derived mesenchymal stem cells‐affinity peptide for tissue engineering
US20130101635A1 (en) Method of preparing self-assembled extracellular matrices and use of self-assembled extracellular matrices formed by using the method
Lee et al. Injectable gel with synthetic collagen-binding peptide for enhanced osteogenesis in vitro and in vivo
Liu et al. Immunomodulatory hybrid micro-nanofiber scaffolds enhance vascular regeneration
US20100151575A1 (en) Method of Making Conditioned Media from Kidney Derived Cells
Niu et al. Comparative study of three types of polymer materials co-cultured with bone marrow mesenchymal stem cells for use as a myocardial patch in cardiomyocyte regeneration
CN109563474B (zh) 整合的细胞
Ghaffarinovin et al. Repair of rat cranial bone defect by using amniotic fluid-derived mesenchymal stem cells in polycaprolactone fibrous scaffolds and platelet-rich plasma
Bai et al. Bioprinted living tissue constructs with layer-specific, growth factor-loaded microspheres for improved enthesis healing of a rotator cuff
Han et al. Sa12b-Modified functional self-assembling peptide hydrogel enhances the biological activity of nucleus pulposus mesenchymal stem cells by inhibiting acid-sensing ion channels
KR101906557B1 (ko) 합성 펩타이드를 이용한 유도만능줄기세포의 제조방법

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11867567

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11867567

Country of ref document: EP

Kind code of ref document: A1