Serum-free culturing system inducing differentiation of pluripotent stem cells into motor neuron cells

A serum-free culture technology for motor nerve cells, which is applied in the fields of stem cell biology and cell culture, can solve the problems of unknown serum components, low repeatability, and complexity, so as to improve safety and application value, reduce differentiation, and reduce batches. The effect of secondary differences

Inactive Publication Date: 2018-12-07
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 ma...

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  • Serum-free culturing system inducing differentiation of pluripotent stem cells into motor neuron 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 motor 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 Smoothened agonist, 2 μM cytarabine.

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

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Abstract

The invention belongs to the technical fields of stem cell biology and cell culturing, and provides a serum-free culturing system inducing differentiation of pluripotent stem cells into motor neuron cells. The serum-free culturing system specially comprises a serum-free differentiation basic culture medium and an additive for promoting the differentiation of the motor neuron cells, wherein necessary components of the serum-free differentiation basic culture medium comprise a DMEM/F12 culture medium, a human serum albumin, L-glutamine, D-glucose, vitamin C, N2/B27 cell culture additive and unnecessary amino acid. The additive for promoting the differentiation of the motor neuron cells specially comprises SB431542, dermomorphin, fibroblast growth factors-2, retinoic acid, BDNF, GDNF, CTNF, DAPT, Smoothened agonist, and cytarabine. The culture medium is adopted to help improve the differentiation of the motor neuron cells in a cell mixing system, the differentiation of other types of cells is reduced, and finally, the motor neuron cells with high purity can be obtained.

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 motor 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, but a...

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Application Information

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IPC IPC(8): C12N5/079
CPCC12N5/0618C12N2500/32C12N2500/34C12N2500/38C12N2500/40C12N2501/115C12N2501/13C12N2501/998
Inventor 甄威刘子铖王楠
Owner 九三极恒生物医药科技江苏有限公司
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