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Method for inducing neuroblastoma cells to be differentiated to nerve cells

A neuroblastoma and neural stem cell technology, applied in the field of cell research, can solve the problems of long differentiation time and low neuron cell differentiation efficiency, and achieve the effect of inhibiting cell apoptosis

Active Publication Date: 2015-12-23
SHANDONG QILU STEM CELL ENG
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Problems solved by technology

[0004] In order to solve the problems of low differentiation efficiency and long differentiation time of neuroblastoma cells differentiated from neuroblastoma cells in the prior art, the present invention provides an induced neuroblastoma cell with a differentiation effect greater than 80% and a short differentiation time Methods of Differentiation into Neurons

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  • Method for inducing neuroblastoma cells to be differentiated to nerve cells
  • Method for inducing neuroblastoma cells to be differentiated to nerve cells
  • Method for inducing neuroblastoma cells to be differentiated to nerve cells

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Embodiment 1

[0024] (1) Preparation of neural stem cell conditioned medium

[0025] Human neural stem cells were purchased from Lonza Company and cultured in T-75 culture flasks with 15 ml of complete medium. Complete medium containing DMEM / F12 basal medium (Invitrogen), human recombinant epidermal growth factor (EGF, 20ng / mL) and basic fibroblast growth factor (bFGF, 20ng / mL) (R&D), B27 (serum replacement , 1:50, Invitrogen), heparin (5 μg / mL, Sigma), L-glutamine 2mM, and penicillin and streptomycin (1:100, Invitrogen). at 37°C with 5% CO 2 Incubate in a humidified incubator, and replace 50% of the medium with new complete medium every 3 days. When the neurospheres grow to a diameter of about 1 mm, they are cut and passaged under a microscope. When the culture medium is replaced each time, the old culture medium replaced is the neural stem cell conditioned medium. The conditioned medium was collected and filtered through a 0.22 μm pore size filter (Millipore), then centrifuged at 1000...

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Abstract

The invention relates to the cell transdifferentiation technology in the field of cell research, in particular to a method for inducing neuroblastoma cells to be differentiated to nerve cells. The neuroblastoma cells are cultured for 3-7 days through an inducing culture medium, and then the nerve cells are obtained; the inducing culture medium is obtained by adding retinoic acid to a neural stem cell condition culture solution to 10 micrometers; a complete culture medium including a DMEM / F12 basic cukture mediam, B27, bFGF, EGF, heparn sodium, L-glutamine and mycillin is used for culturing the neural stem cells, the complete culture medium is replaced by a half after culturing is conducted for three days, and the replaced culture solution is the neural stem cell condition culture solution. By means of the method, the differentiation efficiency is improved, and the maturity of the cells and growth of neural synapses of nerve cells are also improved. The cell apoptosis in SH-SY5Y treated through RA can be inhibited.

Description

technical field [0001] The invention relates to cell transdifferentiation technology in the field of cell research, in particular to a method for inducing neuroblastoma cells to differentiate into neurons. Background technique [0002] It is well known that neurons differentiated from the neuroblastoma SH-SY5Y cell line are commonly used cell models for neurobiological studies, such as neurotoxicity studies and in vitro studies of neuronal tolerance to morphine [Constantinescu, JNeuralTransmSuppl (2007)]. SH-SY5Y is a human-derived cell line that can expand continuously without differentiation under in vitro culture conditions. To differentiate SH-SY5Y cells into neurons, retinoic acid (RA) is usually used for induction. However, the ratio of neuronal cells differentiated from SH-SY5Y cells was very low, whether induced by RA alone or in combination with other compounds. RA alone induced only about 20% of SH-SY5Y cells to differentiate into neurons [Preis, CancerRes (1988)...

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Application Information

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IPC IPC(8): C12N5/0793
Inventor 刘小盾杨洪娜曲廷瑜
Owner SHANDONG QILU STEM CELL ENG
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