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Method for inducing human amniotic mesenchymal stem cells to differentiate into neuron-like cells

A technology of neuron-like cells and mesenchymal stem cells, which can be applied to animal cells, nervous system cells, vertebrate cells, etc., and can solve the problems of lack of physiological characteristics of nerve cells

Inactive Publication Date: 2013-04-03
陆华
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Problems solved by technology

However, most of these studies have focused on the fact that hAMSCs have neuronal morphological characteristics and express neuronal markers after induction, but lack evidence of neuronal physiological characteristics, so many scholars believe that these cells are called "neuron-like cells". more appropriate

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  • Method for inducing human amniotic mesenchymal stem cells to differentiate into neuron-like cells

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[0014] Such as figure 1 Shown is a flow chart of the steps of the method for inducing human amniotic mesenchymal stem cells to differentiate into neuron-like cells of the present invention, which uses all-trans retinoic acid, basic fibroblast growth factor, and epidermal growth factor to induce human amniotic mesenchymal stem cells The method for differentiating stromal stem cells into neuron-like cells specifically comprises the following steps:

[0015] S1. Isolation of human amniotic mesenchymal stem cells (hAMSCs):

[0016] The fresh placenta discarded after delivery was taken under sterile conditions, the amniotic membrane was peeled off from the placental tissue by mechanical method, washed several times with D-Hanks solution to remove residual blood, and the rinsed amniotic membrane was cut into about 1mm with ophthalmic scissors 3 Add 2.5g / L trypsin to digest at 37°C for 10 minutes; add DMEM containing 5% calf serum to stop the digestion, and gently pipette and mix we...

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Abstract

The invention provides a method for inducing differentiating human amniotic mesenchymal stem cells (hAMSCs) to differentiate into neuron-like cells by adopting all-trans retinoic acids, a basic fibroblast growth factor (bFGF) and an epidermal growth factor (EGF). The method comprises the following steps of: separating the hAMSCs, carrying out primary culture of the hAMSCs, subculturing and amplifying the hAMSCs, detecting hAMSCs immunophenotyping, inducing the hAMSCs to differentiate into the neuron-like cells and carrying out cellular immunity fluorescence staining. According to the inducing method provided by the invention, umbilical cord mesenchymal stem cells are induced to differentiate into neutral stem cells by using the all-trans retinoic acids in combination of the bFGF and the EGF; the neural stem cells not only have the typical morphology of nerve cells, but also express neuron marker antigen neuron-specific emolase and astrocyte marker antigen glial fibrillary acidic proteins; and the capability of a mesenchymal cell trans-germinal layer differentiating into non mesenchymal cells is realized so that the mesenchymal cells are likely to turn into more ideal seed cells for clinical application in further.

Description

technical field [0001] The invention relates to a method for inducing human amniotic mesenchymal stem cells (hAMSCs) to differentiate into neuron-like cells in the field of biotechnology, in particular to a method for inducing neuron-like cells using all-trans retinoic acid, basic fibroblast growth factor, and epidermal growth factor A method for differentiating human amniotic mesenchymal stem cells into neuron-like cells. Background technique [0002] Human amnion membrane mesenchymal stem cells (hAMSCs) maintain the plasticity of embryonic stem cells in the early stage of gastrulation and have greater multilineage differentiation potential. hAMSCs can theoretically be differentiated into various tissue cells. At the same time, because of the convenience of obtaining amnion and avoiding ethical disputes, hAMSCs have the potential to become one of the sources of stem cells for future clinical applications. However, the conversion rate of nerve cells after hAMSCs transplanta...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0793C12N5/0775
Inventor 陆华
Owner 陆华
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