Micro RNA capable of inducing stem cell to differentiate toward osteoblasts and use thereof

A kind of adult stem cell and tiny technology, applied in the field of microRNA, can solve the problems of unsustainable curative effect, high toxicity, lack of evidence of cell replacement, etc., and achieve the effect of avoiding poor stability and long half-life

Inactive Publication Date: 2009-03-25
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are several problems in the clinical application of chemically synthesized microRNAs: 1) the stability of microRNAs in body fluids is poor; 2) microRNAs need to be mediated by in vivo transfection reagents to enter cells. The reagents have the problems of poor efficiency, large dosage, and high toxicity; 3) The chemically synthesized microRNA has a short half-life, requires multiple medications, a

Method used

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  • Micro RNA capable of inducing stem cell to differentiate toward osteoblasts and use thereof
  • Micro RNA capable of inducing stem cell to differentiate toward osteoblasts and use thereof
  • Micro RNA capable of inducing stem cell to differentiate toward osteoblasts and use thereof

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Experimental program
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Effect test

Embodiment 1

[0050] Embodiment 1, microRNA induces osteogenic differentiation of stem cells

[0051] 1) Preparation of microRNA

[0052] MicroRNAs are selected from the sequences of mature microRNAs of the human miR-20 family, from miRBase ( http: / / microrna.sanger.ac.uk / sequences / index.shtml ), named respectively as miR-17-5p (its nucleotide sequence is sequence 1 in the sequence listing), miR-18a (its nucleotide sequence is sequence 2 in the sequence listing), miR-18b (its nucleotide sequence The acid sequence is sequence 3 in the sequence listing), miR-20a (its nucleotide sequence is sequence 4 in the sequence listing), miR-20b (its nucleotide sequence is sequence 5 in the sequence listing), miR-93 (its nucleotide sequence is sequence 6 in the sequence listing), miR-106a (its nucleotide sequence is sequence 7 in the sequence listing), miR-106b (its nucleotide sequence is sequence 8 in the sequence listing) and miR-519d (its nucleotide sequence is sequence 9 in the sequence listing). Th...

Embodiment 2

[0084] Embodiment 2, preparation of cells for treating orthopedic diseases

[0085] The patient's own bone marrow was collected, MSCs were collected by Ficoll-Hapague density gradient centrifugation, and then placed in a-MEM medium (purchased from GIBCOL) containing 10g / 100ml fetal bovine serum (FBS) and 1g / 100ml penicillin and streptomycin, 37 °C, 5% CO 2 For culture, the mixed blood cells can be removed through continuous medium replacement and subculture, and pure MSCs can be obtained after 5 passages. Then the MSCs cells were divided into 5×10 5 The density of / hole inoculates 12 orifice plate, after 24 hours, according to the instruction manual of the Lipofectamine2000 of Invitrogen Company, 9 microRNAs (its nucleotide sequence is the sequence 1-9 in the sequence table) and control 1 (its nucleoside) respectively The acid sequence is the sequence 10 in the sequence list) to transfect MSCs cells, and the transfected MSCs cells continue to be cultured for 4-6 days, which...

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Abstract

The invention discloses a micro RNA which can induce differentiation from stem cells to osseous cells and an application thereof. The micro RNA disclosed by the invention is one of the following RNAs, namely a double-stranded RNA of a1) or a single-stranded RNA of b1): a1) the nucleotide sequence of one strand is a sequence 1-9 in a sequence table while that of the other strand is a reverse complement to the sequence 1-9 in the sequence table; b1) a recombinant RNA molecule is obtained by connecting a single-stranded RNA with the nucleotide sequence which is the sequence 1-9 in the sequence table and a single-stranded RNA with the nucleotide sequence which is a reverse complement to the sequence 1-9 in the sequence table by a connection sequence; and the connection sequence is TTCAAGAGA. The micro RNA can be used for preparation of medicaments for the treatment of orthopedic disorders and the medicaments can be adopted for the treatment of the orthopedic disorders of osteoporosis, bone fracture, hairline fracture, femoral head necrosis and the like.

Description

technical field [0001] The present invention relates to microRNA capable of inducing stem cells to differentiate into osteoblasts and application thereof. Background technique [0002] MicroRNA (microRNA) is a kind of non-coding RNA with a length of about 21-24 nucleotides, which plays an important role in the regulation of gene expression. MicroRNA forms a complex with Ago and other enzymes and proteins to bind to the 3' untranslated region of mRNA of the target gene, hindering the translation of mRNA or causing mRNA degradation. Evidence suggests that microRNAs are involved in a series of important processes in the life process, and have biological functions such as regulating cell proliferation, differentiation, death, and individual development. MicroRNAs can be regarded as a class of endogenous small interfering RNAs. Small RNA will become the third generation of drugs after chemical drugs and protein drugs. However, there are several problems in the clinical applica...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N5/10A61K31/713A61K31/7088A61K48/00A61L27/38A61F2/82A61P19/10A61P19/08C12N15/113
Inventor 张雅鸥孔祥復胡祥李国喜张锦芳谢伟东
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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