Serum-free culture system capable of promoting induced pluripotent stem cells to be differentiated to cortical nerve cells

A serum-free culture, pluripotent stem cell technology, applied in the field of serum-free culture system, can solve the problems of low repeatability, unknown serum composition, complexity, etc., to reduce batch differences, improve safety and application value, reduce The effect of differentiation

Inactive Publication Date: 2018-11-27
九三极恒生物医药科技江苏有限公司
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Problems solved by technology

[0004] Most of the medium for pluripotent stem cell culture and differentiation into nerve cells contains fetal bovine serum components. The components of serum are unknown and complex. Due to the large differences in batches of serum products from different manuf

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  • Serum-free culture system capable of promoting induced pluripotent stem cells to be differentiated to cortical nerve cells

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Embodiment Construction

[0013] The present invention will be described in more detail below in conjunction with the drawings and embodiments, but should not be considered limited to the embodiments described herein.

[0014] DMEM / F12 was used as the basal medium, and the following components were added to it to prepare a serum-free basal medium: 2.5g / L serum albumin, 5nM L-glutamine, 2g / L D-glucose, 200μg / L Vitamin C in L, 1% N2 cell culture supplement, 2% B27 cell culture supplement, 100 μM non-essential amino acids.

[0015] Additives to promote the differentiation of cortical nerve cells, the specific component concentrations are: 10 μM SB431542, 1 μM dermorphin, 10 μg / L fibroblast growth factor-2, 1 μM retinoic acid, 1 μg / L brain-derived neurotrophic factor , 1 μg / L glial cell-derived neurotrophic factor, 1 μg / L ciliary ganglion neurotrophic factor, 2 μM DAPT, 2 μM cytarabine.

[0016] Human induced pluripotent stem cells were cultured in a serum-free minimal medium system to form embryoid bodie...

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Abstract

The invention belongs to the technical field of stem cell biology and cell culture and provides a serum-free culture system capable of promoting induced pluripotent stem cells to be differentiated tocortical nerve cells. The serum-free culture system specifically comprises a serum-free differentiation basic culture medium and additives capable of promoting cortical nerve cell differentiation, andnecessary ingredients of the serum-free basic culture medium include DMEM/F12 culture medium, human serum albumin, L-glutamine, D-glucose, vitamin C, N2/B27 cell culture additive and non-essential amino acid. The additives capable of promoting cortical nerve cell differentiation specifically include SB431542, dermorphin, fibroblast growth factor-2, retinoic acid, BDNF, GDNF, CTNF, DAPT and cytarabine. By using the culture medium, cortical nerve cell differentiation in a cell mixed system can be promoted, and differentiation towards cells of other types is reduced, so that high-purity corticalnerve cells are obtained finally.

Description

technical field [0001] The invention belongs to the technical fields of stem cell biology and cell culture, and specifically relates to a serum-free culture system for promoting the differentiation of induced pluripotent stem cells into cortical nerve cells. Background technique [0002] Neurodegenerative diseases are a kind of diseases with slow onset, progressive development and poor prognosis. So far, there is no effective cure method. Common clinical neurodegenerative diseases include Parkinson's disease, amyotrophic lateral sclerosis, spinal muscular atrophy, and Alzheimer's disease. Although these diseases have different etiologies, they all have different parts of the central nervous system and different degrees of neuron loss and dysfunction in pathology. Replacing lost neurons and restoring their function is a new approach to treating these diseases. Studies in recent years have found that stem cells cultured in vitro can not only proliferate and differentiate, bu...

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Application Information

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IPC IPC(8): C12N5/079
CPCC12N5/0618C12N2500/32C12N2500/34C12N2500/38C12N2500/40C12N2500/90C12N2501/115C12N2501/13C12N2501/15C12N2501/734C12N2501/85C12N2501/998C12N2506/45
Inventor 甄威刘子铖康健周海荣
Owner 九三极恒生物医药科技江苏有限公司
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