CN112342217A - Periodontal ligament stem cell proliferation and osteogenic differentiation promoter - Google Patents
Periodontal ligament stem cell proliferation and osteogenic differentiation promoter Download PDFInfo
- Publication number
- CN112342217A CN112342217A CN202011252094.3A CN202011252094A CN112342217A CN 112342217 A CN112342217 A CN 112342217A CN 202011252094 A CN202011252094 A CN 202011252094A CN 112342217 A CN112342217 A CN 112342217A
- Authority
- CN
- China
- Prior art keywords
- shrna
- periodontal ligament
- ligament stem
- osteogenic differentiation
- sequence
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
- C07K14/4705—Regulators; Modulating activity stimulating, promoting or activating activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0654—Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/999—Small molecules not provided for elsewhere
Abstract
The invention provides a periodontal ligament stem cell proliferation and osteogenic differentiation promoter, belonging to the technical field of stem cells. The invention firstly discovers that the expression level of the non-coding RNA RP11-289F5.1 is reduced in the process of osteogenic differentiation of periodontal ligament stem cells. Secondly, shRNA for silencing RP11-289F5.1 is designed and synthesized, and the silencing RP11-289F5.1 is proved to be capable of effectively promoting osteogenic differentiation and proliferation of periodontal ligament stem cells.
Description
Technical Field
The invention belongs to the technical field of periodontal ligament stem cells, and particularly relates to a periodontal stem cell proliferation and osteogenic differentiation promoter.
Background
Periodontitis is one of the common oral diseases, a chronic inflammatory disease of the periodontal supporting tissue. Clinically, periodontitis patients are characterized primarily by: probing bleeding, gingival swelling, periodontal formation and alveolar bone resorption, and finally tooth loosening and falling off. At present, the conventional methods for treating periodontitis include basic treatment such as periodontal scaling and scraping, periodontal surgery treatment such as periodontal flap surgery and root planing surgery, and combined photodynamic treatment. However, these methods only provide temporary relief to the patient and do not provide a substantial recovery of the periodontal support tissue.
The periodontal ligament stem cell is a stem cell separated from the periodontal ligament and has strong self-proliferation and multi-directional differentiation capacity. Periodontal ligament stem cells can be induced under certain conditions to reprogram them to achieve the purpose of lateral differentiation, and thus become seed cells for tissue regeneration engineering. Therefore, it is of great significance to promote the osteogenic differentiation and proliferation ability of periodontal ligament stem cells.
Disclosure of Invention
The purpose of the present invention is to provide an agent for promoting the proliferation and osteogenic differentiation of periodontal ligament stem cells.
In order to achieve the purpose, the invention provides the following technical scheme:
in a first aspect of the invention, a gene for promoting proliferation of periodontal ligament stem cells is RP11-289F5.1, the transcription sequence of the gene is ENST00000397864.2, and the sequence of ENST00000397864.2 is shown in SEQ ID NO. 1.
In a second aspect of the invention, the application of the RP11-289F5.1 silencing agent in preparing the periodontal ligament stem cell proliferation promoter is provided.
Preferably, the silencing agent is selected from shRNA, siRNA, small molecule compounds.
Preferably, the silencing reagent is shRNA, the sequence of the sense strand of the shRNA is shown as SEQ ID NO.6, and the sequence of the antisense strand of the shRNA is shown as SEQ ID NO. 7.
In a third aspect of the invention, the promoter is shRNA of RP11-289F5.1, the sense strand sequence of the shRNA is shown as SEQ ID NO.6, and the antisense strand sequence of the shRNA is shown as SEQ ID NO. 7.
In a fourth aspect of the invention, the invention provides an application of an RP11-289F5.1 silencing reagent in preparing a periodontal ligament stem cell osteogenic differentiation promoter.
Preferably, the silencing agent is selected from shRNA, siRNA, small molecule compounds.
Preferably, the silencing reagent is shRNA, the sequence of the sense strand of the shRNA is shown as SEQ ID NO.6, and the sequence of the antisense strand of the shRNA is shown as SEQ ID NO. 7.
The fifth aspect of the invention provides an accelerant for promoting osteogenic differentiation of periodontal ligament stem cells, wherein the accelerant is shRNA of RP11-289F5.1, the sense strand sequence of the shRNA is shown as SEQ ID NO.6, and the antisense strand sequence of the shRNA is shown as SEQ ID NO. 7.
The invention has the beneficial effects that the expression quantity of the non-coding RNA RP11-289F5.1 is down-regulated in the process of osteogenic differentiation of periodontal ligament stem cells for the first time. Secondly, shRNA for silencing RP11-289F5.1 is designed and synthesized, and the silencing RP11-289F5.1 is proved to be capable of effectively promoting osteogenic differentiation and proliferation of periodontal ligament stem cells.
Drawings
FIG. 1 shows the change of the expression level of RP11-289F5.1 during the osteogenic differentiation of periodontal ligament stem cells.
FIG. 2 shows the results of the test of the inhibition effect of sh-RP11-289F5.1 designed by the invention.
FIG. 3 silencing regulation of ALP enzymes by RP11-289F 5.1.
FIG. 4 silencing regulation of cell mineralization by RP11-289F 5.1.
FIG. 5 silencing regulation of RP11-289F5.1 on osteogenic differentiation-related proteins RUNX2 and OCN.
FIG. 6 silencing regulation of proliferation of periodontal ligament stem cells by RP11-289F 5.1.
Detailed Description
In order to clearly illustrate the technical features of the present solution, the present solution is explained below by way of specific embodiments.
Detection of the change of RP11-289F5.1 in the osteogenic differentiation of periodontal ligament Stem cells
1. Cell processing
(1) Experiments are divided into two groups, a control group is cultured by using a common complete culture medium, and an experimental group is cultured by using an osteogenic induction culture medium (100 uM vitamin C, 10nM beta-sodium glycerophosphate and 10nM dexamethasone are added into an alpha-MEM complete culture medium containing 10% FBS);
(2) inoculating primary cultured periodontal ligament stem cells into a 6-well cell culture plate, culturing according to cell groups for 7 days, and detecting the expression level of RP11-289F5.1 by using fluorescent quantitative PCR (polymerase chain reaction) on 1,3,5 and 7 days respectively;
2. fluorescent quantitative PCR detection
(1) Washing the treated cells by PBS, adding 1ml of Trizol reagent into each hole, and extracting total RNA of the periodontal ligament stem cells according to the instructions of the Trizol reagent;
(2) reverse transcribing RNA to cDNA according to the instructions of the genomic DNA reverse transcription kit, and storing at-20 ℃;
(3) RP11-289F5.1 and GAPDH primers were designed with the following primer sequences:
(4) a fluorescent quantitative PCR reaction solution was prepared as follows
(5) And (3) PCR reaction conditions:
stage1 pre-deformed Reps: 195 ℃ for 30 s; stage 2: PCR reaction Reps: 4095 deg.C, 5s, 60 deg.C, 30 s;
(6) use 2-ΔΔCtThe method performs data analysis, and the results are shown in FIG. 1.
From the experimental results, it can be seen that the expression level of RP11-289F5.1 gradually decreased during the osteogenic differentiation of periodontal ligament stem cells, and at the 7 th day of culture, the expression level of RP11-289F5.1 in the control group was 0.973 + -0.009, and the expression level of RP11-289F5.1 in the experimental group was 0.492 + -0.018, the difference being statistically significant. As described above, the expression level of RP11-289F5.1 was gradually decreased during the osteogenic differentiation of periodontal ligament stem cells.
Example 2
Silencing RP11-289F5.1 promotes ALP enzyme activity of dental pulp stem cells
Since the expression of RP11-289F5.1 is gradually reduced in the osteogenic differentiation process of the dental pulp stem cells, we suspect that inhibiting the expression of RP11-289F5.1 can effectively promote the osteogenic differentiation of the dental pulp stem cells.
1. sh-RP11-289F5.1 was designed and inhibitory effects were verified
(1) The shRNA of sh-RP11-289F5.1, a shRNA of synthetic RP11-289F5.1, was designed as follows (using the vector pENTR ™ U6):
| sh-RP11-289F5.1 | sequence of |
| Top Strand | 5'-CACCGCTGAAGAACGCAATCCAACAGAGATGTTGGATTGCGTTCTTCAGC -3'，SEQ ID NO.6 |
| Bottom Strand | 5'-AAAAGCTGAAGAACGCAATCCAACATCTCTGTTGGATTGCGTTCTTCAGC -3',SEQ ID NO.7 |
(2) The inhibition effect of sh-RP11-289F5.1 was detected by fluorescent quantitative PCR, and the experimental results are shown in FIG. 2. As can be seen from the figure, the expression level of RP11-289F5.1 of the group sh-RP11-289F5.1 is 0.263 +/-0.046, which indicates that the sh-RP11-289F5.1 can effectively inhibit RP11-289F 5.1.
2. Silencing RP11-289F5.1 Effect on ALP enzymatic Activity of periodontal Membrane Stem cells
(1) The experiment was divided into 3 groups, and the NC group was cultured using complete medium; the sh-NC group is transfected with sh-NC and cultured by using an osteogenic differentiation culture medium; group sh-RP11-289F5.1 transfects RP11-289F5.1 and is cultured by using osteogenic differentiation medium;
(2) culturing according to the experimental groups of the step (1), after osteogenic induced differentiation for 7 days, washing cells by using PBS (phosphate buffer solution), rinsing the cells for 2 times, and fixing the cells by using 4% paraformaldehyde for 30 min;
(3) discarding the fixing solution, rinsing with PBS for 2 times, preparing BCIP/NBT staining working solution, adding 1.5ml of BCIP/NBT staining solution into each hole, and incubating for 30min at room temperature in a dark place;
(4) the BCIP/NBT staining solution was removed, rinsed 2 times with deionized water, and recorded by photography.
The experimental results are shown in FIG. 3, and it can be seen that there is no significant ALP staining in NC group, ALP staining is darker in sh-RP11-289F5.1 than in sh-NC group, which indicates that silencing RP11-289F5.1 can effectively promote the activity of ALP enzyme of periodontal ligament stem cell.
Example 3
Silencing RP11-289F5.1 Studies its effects on mineralization of periodontal ligament stem cells
(1) The experiment was divided into 3 groups, and the NC group was cultured using complete medium; the sh-NC group is transfected with sh-NC and cultured by using an osteogenic differentiation culture medium; group sh-RP11-289F5.1 transfects RP11-289F5.1 and is cultured by using osteogenic differentiation medium;
(2) culturing according to the experimental groups of the step (1), after osteogenic induced differentiation is carried out for 21 days, washing cells by using PBS (phosphate buffer solution), rinsing the cells for 2 times, and fixing the cells by using 4% paraformaldehyde for 30 min;
(3) discarding the stationary liquid, rinsing with PBS for 2 times, preparing alizarin red staining solution, adding 1.5ml of alizarin red staining solution into each hole, and incubating for 30min at room temperature in a dark place;
(4) and removing alizarin red dye liquor, rinsing with deionized water for 2 times, and photographing and recording.
The experimental result is shown in figure 4, and as can be seen from the figure, the NC group has no obvious mineralized nodule formation, and the mineralization degree of the sh-RP11-289F5.1 group is obviously higher than that of the sh-NC group, which indicates that the silent RP11-289F5.1 can promote the mineralization level of periodontal ligament stem cells.
Example 4
(1) The experiment was divided into 3 groups, and the NC group was cultured using complete medium; the sh-NC group is transfected with sh-NC and cultured by using an osteogenic differentiation culture medium; group sh-RP11-289F5.1 transfects RP11-289F5.1 and is cultured by using osteogenic differentiation medium;
(2) after osteogenic differentiation induction for 14 days, the medium was removed and washed 3 times with PBS;
(3) adding cell lysis solution to lyse cells, collecting lysis solution to 1.5ml EP tube, ultrasonically crushing to extract protein for 5min, and suspending circulation at 2s ultrasound/3 s;
(4) centrifuging at 4 ℃ at 12000r/min for 20min, and collecting precipitate;
(5) protein concentration was measured using BCA method, 5 x SDS loading buffer was added;
(6) preparing 12% separation gel, loading 20ug, concentrating gel at 80V for 30min, and allowing 120V separation gel to reach the bottom of separation gel;
(7) after electrophoresis is finished, sequentially placing a spongy cushion, filter paper gel, a PVDF (polyvinylidene fluoride) membrane, filter paper and a spongy cushion on a black surface of an electric rotating clamp, removing bubbles and fastening, and rotating for 1.5 hours at a constant current of 250 mA;
(8) after the electrotransformation is finished, taking out the PVDF membrane, rinsing the PVDF membrane for 5min by using TBST, then adding the PVDF membrane into 5% skimmed milk powder, and sealing the PVDF membrane for 1h by a shaking table at room temperature;
(9) cutting the membrane according to the size of RUNX2, OCN and beta-actin protein, putting the membrane into a small groove with an antibody, and incubating overnight at 4 ℃;
(10) taking out the PVDF membrane, and washing the PVDF membrane in TBST for 3 times, 10min each time;
(11) incubating the secondary antibody according to the requirements of the corresponding antibody, and incubating for 1h at room temperature;
(12) the secondary antibody was removed, and the film was washed 3 times with TBST for 10min each time, and then developed and exposed by adding a chemiluminescent solution.
The experimental results are shown in figure 5, and it can be seen from the figure that the expression levels of RUNX2 and OCN protein of the sh-RP11-289F5.1 group are up-regulated compared with the sh-NC group, which indicates that the silencing of RP11-289F5.1 can effectively promote the expression of the periodontal ligament stem cell osteogenic differentiation related proteins RUNX2 protein and OCN protein.
The combination of examples 2-4 shows that silencing RP11-289F5.1 can effectively promote osteogenic differentiation of periodontal ligament stem cells.
Example 5
(1) The experimental grouping is sh-NC group and sh-RP11-289F5.1 group, the sh-NC group transfects sh-NC, the sh-RP11-289F5.1 group transfects sh-RP11-289F 5.1;
(2) inoculating periodontal ligament stem cells of third generation logarithmic growth stage in 96-well plate, adding 2000 cells per well, continuously culturing for 7 days, and changing liquid 1 time every two days;
(3) the absorbance was measured at 1,3,5, and 7 days using CCK-8 and a growth curve was plotted.
As can be seen from the figure, the proliferation rate of the sh-RP11-289F5.1 group is significantly faster than that of the sh-NC group. Wherein, the difference between the two is not statistically significant at 1 day; at 3 days, the OD value of the sh-NC group is 0.260 +/-0.029, the OD value of the sh-RP11-289F5.1 group is 0.399 +/-0.041, and P is less than 0.05; at 5 days, the OD value of the sh-NC group is 0.376 +/-0.043, the OD value of the sh-RP11-289F5.1 group is 0.516 +/-0.042, and the P is less than 0.05; at 7 days, the OD value of the sh-NC group is 0.436 +/-0.43, the OD value of the sh-RP11-289F5.1 group is 0.606 +/-0.042, and the P is less than 0.05.
The results show that the silencing of RP11-289F5.1 can effectively promote the proliferation of periodontal ligament stem cells.
The technical features of the present invention that are not described can be implemented by or using the prior art, and are not described herein again, of course, the above description is not intended to limit the present invention, and the present invention is not limited to the above examples, and variations or substitutions made by those skilled in the art within the spirit scope of the present invention should also fall into the protection scope of the present invention.
Sequence listing
<110> Yanggui plum
<120> an agent for promoting the proliferation and osteogenic differentiation of periodontal ligament stem cells
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 791
<212> DNA
<213> Human source (Human)
<400> 1
gacaggatcc cactgtgtca cccaggctga agtgcagtgg caccattata gctcactgca 60
gccttgaact cttgggctca agtgatcctc ctacctcagc ctcccgagta gctgggacta 120
cagaaatgtc aattgcctac ctgatggctg tgaaaaacag aaagaagaaa caaagacata 180
tctgtatgta aaggttgagc ttgaaaaaag aaaactactc ttgacgctga agaacgcaat 240
ccaacatgaa agaatgctta tttcaagttt tacctgaatc tacctagagc tgaatttctt 300
atctatactt tactacagat attctgcttt gtgcagatct acaatggatc attaaagtca 360
aaggacaatt acttatcttt ctactcctcc cagtacccaa gacctgagag tgagatatta 420
aattcgtcct tagatctttc atgcaatgtt acagctcttt ggaactcctc atgaagctcc 480
tcctctccat ggtggaattt tactctgagg agagataggc taaaatgttt cagtccccta 540
tatgataagt ccaaaatcac cgtttcacat atatgctgac ctgatacatt gagtcactga 600
tacgtgtctt tccaccttca gtatattttc tttgactagc tttattgctt ccctgtgatg 660
ctgaattttg agcatggatg tgtgtgtcca taacaaaacc atcctaggta aactgaattt 720
ggttatttga atttcataat tactctttac tccaagtgtt gtggattcat gatgcaaatt 780
tacccattcc c 791
<210> 2
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
ctactcctcc cagtacccaa ga 22
<210> 3
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
ggagcttcat gaggagttcc aa 22
<210> 4
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
ggtcaccagg gctgctttta 20
<210> 5
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
ggatctcgct cctggaagat g 21
<210> 6
<211> 50
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 6
caccgctgaa gaacgcaatc caacagagat gttggattgc gttcttcagc 50
<210> 7
<211> 50
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
aaaagctgaa gaacgcaatc caacatctct gttggattgc gttcttcagc 50
Claims (9)
1. The gene for promoting the proliferation of periodontal ligament stem cells is RP11-289F5.1, the transcription sequence of the gene is ENST00000397864.2, and the sequence of the ENST00000397864.2 is shown as SEQ ID NO. 1.
Use of RP11-289F5.1 silencing agent in preparation of periodontal ligament stem cell proliferation promoter.
3. The use according to claim 2, wherein the silencing agent is selected from the group consisting of shRNA, siRNA, and small molecule compounds.
4. The use according to claim 3, wherein the silencing agent is an shRNA, the sequence of the sense strand of the shRNA is shown as SEQ ID No.6, and the sequence of the antisense strand of the shRNA is shown as SEQ ID No. 7.
5. The promoter for promoting the proliferation of the periodontal ligament stem cells is shRNA of RP11-289F5.1, the sense strand sequence of the shRNA is shown as SEQ ID NO.6, and the antisense strand sequence of the shRNA is shown as SEQ ID NO. 7.
6. Application of RP11-289F5.1 silencing reagent in preparation of periodontal ligament stem cell osteogenic differentiation promoter.
7. The use of claim 6, wherein the silencing agent is selected from the group consisting of shRNA, siRNA, and small molecule compounds.
8. The use according to claim 7, wherein the silencing agent is an shRNA, the sequence of the sense strand of the shRNA is shown as SEQ ID No.6, and the sequence of the antisense strand of the shRNA is shown as SEQ ID No. 7.
9. The promoter for promoting osteogenic differentiation of periodontal ligament stem cells is shRNA of RP11-289F5.1, wherein the sense strand sequence of the shRNA is shown as SEQ ID NO.6, and the antisense strand sequence of the shRNA is shown as SEQ ID NO. 7.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110427545.0A CN113106097B (en) | 2020-11-11 | 2020-11-11 | Promoter for osteogenic differentiation of periodontal ligament stem cells |
| CN202011252094.3A CN112342217B (en) | 2020-11-11 | 2020-11-11 | Periodontal ligament stem cell proliferation and osteogenic differentiation promoter |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011252094.3A CN112342217B (en) | 2020-11-11 | 2020-11-11 | Periodontal ligament stem cell proliferation and osteogenic differentiation promoter |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110427545.0A Division CN113106097B (en) | 2020-11-11 | 2020-11-11 | Promoter for osteogenic differentiation of periodontal ligament stem cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112342217A true CN112342217A (en) | 2021-02-09 |
| CN112342217B CN112342217B (en) | 2021-04-16 |
Family
ID=74363330
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011252094.3A Active CN112342217B (en) | 2020-11-11 | 2020-11-11 | Periodontal ligament stem cell proliferation and osteogenic differentiation promoter |
| CN202110427545.0A Active CN113106097B (en) | 2020-11-11 | 2020-11-11 | Promoter for osteogenic differentiation of periodontal ligament stem cells |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110427545.0A Active CN113106097B (en) | 2020-11-11 | 2020-11-11 | Promoter for osteogenic differentiation of periodontal ligament stem cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN112342217B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113106097A (en) * | 2020-11-11 | 2021-07-13 | 青岛思拓新源细胞医学有限公司 | Promoter for osteogenic differentiation of periodontal ligament stem cells |
| CN114107184A (en) * | 2021-11-26 | 2022-03-01 | 山东中医药大学第二附属医院 | Application of siRNA in osteogenic differentiation of dental pulp stem cells |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150196371A1 (en) * | 2014-01-13 | 2015-07-16 | Brock B. WESTOVER | Endosseous dental implant assembly |
| CN107354127A (en) * | 2017-07-11 | 2017-11-17 | 山东大学 | Effects of the LncRNA TUG1 in regulation and control PDLSCs Osteoblast Differentiations and regeneration |
| CN110241196A (en) * | 2019-05-10 | 2019-09-17 | 山东大学 | Application of the circRNA PRKD3 in periodontal ligament stem cell Osteoblast Differentiation |
| CN110616190A (en) * | 2019-02-19 | 2019-12-27 | 中国人民解放军第四军医大学 | Method for regulating and controlling osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111235097B (en) * | 2020-03-18 | 2020-10-23 | 山东殷氏干细胞工程有限责任公司 | Application of LINC01877 in osteogenic differentiation of bone marrow mesenchymal stem cells |
| CN111454953B (en) * | 2020-04-16 | 2021-11-26 | 赛尔斯(山东)生物工程有限公司 | Bone marrow mesenchymal stem cell adipogenic transformation promoter |
| CN111658775B (en) * | 2020-06-22 | 2022-08-12 | 北京中卫医正科技有限公司 | Role of long-chain non-coding RNA in osteogenic differentiation and adipogenic differentiation of stem cells |
| CN111534487B (en) * | 2020-06-22 | 2021-05-14 | 孙德强 | Promoter for promoting osteogenic differentiation of mesenchymal stem cells |
| CN112342217B (en) * | 2020-11-11 | 2021-04-16 | 杨桂梅 | Periodontal ligament stem cell proliferation and osteogenic differentiation promoter |
-
2020
- 2020-11-11 CN CN202011252094.3A patent/CN112342217B/en active Active
- 2020-11-11 CN CN202110427545.0A patent/CN113106097B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150196371A1 (en) * | 2014-01-13 | 2015-07-16 | Brock B. WESTOVER | Endosseous dental implant assembly |
| CN107354127A (en) * | 2017-07-11 | 2017-11-17 | 山东大学 | Effects of the LncRNA TUG1 in regulation and control PDLSCs Osteoblast Differentiations and regeneration |
| CN110616190A (en) * | 2019-02-19 | 2019-12-27 | 中国人民解放军第四军医大学 | Method for regulating and controlling osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix |
| CN110241196A (en) * | 2019-05-10 | 2019-09-17 | 山东大学 | Application of the circRNA PRKD3 in periodontal ligament stem cell Osteoblast Differentiation |
Non-Patent Citations (5)
| Title |
|---|
| ENSEMBLE: "Transcript: AL138749.1-201", 《ENSEMBL》 * |
| QIAN QU等: "Potential Role of Long Non-Coding RNA in Osteogenic Differentiation of Human Periodontal Ligament Stem Cells", 《J PERIODONTOL》 * |
| TRACEY,A: "Human DNA sequence from clone RP11-289F5 on chromosome 9q21.31-22.1, complete", 《GENBANK》 * |
| YUERONG XU等: "LncRNA-TWIST1 Promoted Osteogenic Differentiation Both in PPDLSCs and in HPDLSCs by Inhibiting TWIST1 Expression", 《BIOMED RESEARCH INTERNATIONAL》 * |
| 马灵芝等: "miR-21对人牙周膜干细胞增殖及成骨分化的影响", 《口腔疾病防治》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113106097A (en) * | 2020-11-11 | 2021-07-13 | 青岛思拓新源细胞医学有限公司 | Promoter for osteogenic differentiation of periodontal ligament stem cells |
| CN114107184A (en) * | 2021-11-26 | 2022-03-01 | 山东中医药大学第二附属医院 | Application of siRNA in osteogenic differentiation of dental pulp stem cells |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113106097B (en) | 2022-08-02 |
| CN113106097A (en) | 2021-07-13 |
| CN112342217B (en) | 2021-04-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112342217B (en) | Periodontal ligament stem cell proliferation and osteogenic differentiation promoter | |
| CN111454953B (en) | Bone marrow mesenchymal stem cell adipogenic transformation promoter | |
| CN111235097B (en) | Application of LINC01877 in osteogenic differentiation of bone marrow mesenchymal stem cells | |
| CN111658775B (en) | Role of long-chain non-coding RNA in osteogenic differentiation and adipogenic differentiation of stem cells | |
| CN112301032B (en) | Gene inhibitor for inhibiting hair follicle stem cell aging | |
| CN111118013B (en) | Leukemia biomarker LINC02033 and application thereof | |
| CN111500578A (en) | Circ RNA-FTO for regulating and controlling osteogenic differentiation and tissue regeneration of ADSCs and application thereof | |
| Lopatina et al. | The angiogenic potential of adipose mesenchymal stem cell-derived extracellular vesicles is modulated by basic fibroblast growth factor | |
| CN110894488A (en) | Application of all-trans retinoic acid in mesenchymal stem cell differentiation regulation | |
| CN113151177B (en) | Breast or breast cancer tissue acellular matrix and preparation method and application thereof | |
| CN115851972B (en) | Sheep hair follicle development marker miR-23b and application thereof | |
| CN106350519A (en) | MicroRNA for adjusting rapid growth of cornua cervi pantotrichum cartilage and application thereof | |
| CN114292850B (en) | circRNA related to proliferation and differentiation of goat myoblasts and application thereof | |
| CN116355898A (en) | Application of miRNA-133 in regulation of sheep embryo hair follicle development | |
| CN113151267B (en) | Gene inhibitor for reducing oxidative stress apoptosis of umbilical cord blood mesenchymal stem cells | |
| CN105688227B (en) | Application of miR-127 in preparation of medicine for treating muscle diseases | |
| CN114107460A (en) | Application of gene inhibitor in inhibition of aging of umbilical cord blood mesenchymal stem cells | |
| CN113694076A (en) | Application of miRNA and human mesenchymal stem cells in preparation of medicine for treating arthritis | |
| CN113549593B (en) | Pharmaceutical preparation for promoting proliferation of epidermal stem cells | |
| CN113350368B (en) | Application of gene inhibitor in preparation of epidermal stem cell migration pharmaceutical preparation | |
| CN111454892A (en) | Tooth mesenchymal stem cell culture medium and activity verification method in dental pulp stem cells | |
| CN114657121B (en) | Application of LOX1 gene as BMSCs osteogenic differentiation promoter under action of fluid shear force | |
| CN110904038B (en) | Mesenchymal stem cells and application thereof | |
| CN111118154B (en) | Application of LINC01272 in preparation of tumor detection reagent and/or treatment drug | |
| CN108300763A (en) | A method of screening miR-101-3P target genes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220125 Address after: 250000 No. eight, No. 2, Shizhong Road, Shizhong District, Shandong, Ji'nan Patentee after: THE SECOND AFFILIATED HOSPITAL SHANDONG University OF TRADITIONAL CHINESE MEDICINE Address before: No.2, Jingba Road, Shizhong District, Jinan City, Shandong Province Patentee before: Yang Guimei |
|
| CP02 | Change in the address of a patent holder | ||
| CP02 | Change in the address of a patent holder |
Address after: 250000 No. eight, No. 1, Shizhong Road, Shizhong District, Shandong, Ji'nan Patentee after: THE SECOND AFFILIATED HOSPITAL SHANDONG University OF TRADITIONAL CHINESE MEDICINE Address before: 250000 No. eight, No. 2, Shizhong Road, Shizhong District, Shandong, Ji'nan Patentee before: THE SECOND AFFILIATED HOSPITAL SHANDONG University OF TRADITIONAL CHINESE MEDICINE |