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Method for obtaining inner ear hair cell precursor induced by bone marrow mesenchymal stem cells

A technology of bone marrow mesenchyme and inner ear hair cells, applied in artificial cell constructs, bone/connective tissue cells, animal cells, etc., to achieve significant clinical effects

Inactive Publication Date: 2008-07-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on cultured bone marrow MSC directed to differentiate into cochlear sensory epithelial cells in vitro, the key is to find the appropriate induction factor

Method used

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  • Method for obtaining inner ear hair cell precursor induced by bone marrow mesenchymal stem cells
  • Method for obtaining inner ear hair cell precursor induced by bone marrow mesenchymal stem cells
  • Method for obtaining inner ear hair cell precursor induced by bone marrow mesenchymal stem cells

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Experimental program
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Embodiment 1

[0034] 1. Materials and methods

[0035] 1.1 Experimental materials

[0036] 1.1.1 Experimental animals: SD rats, 4-5 weeks old, weighing 80-120 g, were purchased from the Experimental Animal Center of Zhejiang University.

[0037] 1.1.2 Experimental reagents

[0038] 1.1.2.1 General reagents

[0039] 25cm 2 Culture flasks Orange scientific Belgium

[0040] 6-well culture plate Becton Dickinson, USA

[0041] PBS solution Gino Biomedical Technology Co., Ltd. China

[0042] 0.25% Trypsin GIBCO Corporation USA

[0043] Polylysine solution SIGMA Corporation USA

[0044] Premium Fetal Bovine Serum FBS Hangzhou Sijiqing Biological Engineering Co., Ltd. China

[0045] Low sugar DMEM solution GIBCO company USA

[0046] N2 adjuvant GIBCO / invitrogen USA

[0047] 2.1.2.2 Cell induction reagents

[0048] ATRA SIGMA Corporation USA

[0049] EGF CYTOLAB / PEPROTECH USA

[0050] bFGF CYTOLAB / PEPROTECH USA

[0051] IGF-1 CYTOLAB / PEPROTECH USA

[0052] BDNF CYTOLAB / PEPROTECH USA ...

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Abstract

The invention provides a method for inducing mesenchymal stem cell in the bone marrow to obtain the inner ear hair cell prosoma, which comprises the following steps: doing generation culture for the separated and purified mesenchymal stem cell in the bone marrow, switching stable generation mesenchymal stem cell into induction culture medium, inducing and culturing for 7-20 d, obtaining the inner ear hair cell prosoma. The induction culture medium is basic culture medium which is charged with growth factor, wherein the growth factor mainly comprises an epiderm growth factor (EGF) and an insulin growth factor-1 (IGF-1), the charging quantity is EGF 10-30ng / ml basic culture medium and IGF-1 40-60ng / ml basic culture medium. The beneficial effect of the invention is that the invention provides a method for inducing mesenchymal stem cell in the bone marrow to obtain the inner ear hair cell prosoma and supplies a firm base for cochlear cell transplantation therapeutic sound perception nerve deafness, which is provided with significant clinical meaning.

Description

(1) Technical field [0001] The invention relates to a method for inducing inner ear hair cell precursors by using bone marrow mesenchymal stem cells. (2) Background technology [0002] Sensorineural hearing loss is very common clinically, and the latest report reaches billions of people, accounting for about 10% of the total population. There are more than 900 million people in Europe and the United States. Mainly because the number of hair cells in the inner ear is limited, only about 1500; vulnerable to damage or destruction by aging, noise, ototoxic drugs, trauma, infection, tumors, genetic diseases, immune diseases and other internal and external factors; and breastfeeding Animal cochlear hair cells have limited regenerative ability after destruction, and often cause permanent hearing loss, which is very difficult to treat. With the deepening of stem cell research, compensating for the loss of hair cells through stem cell transplantation or regeneration is one of the m...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/08C12N5/071C12N5/0775
Inventor 徐娅苹管明雷云秋彭佳萍
Owner ZHEJIANG UNIV
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