Serum-free culture system for inducing differentiation of multipotent stem cells into neural progenitor cells

A serum-free culture technology of neural progenitor cells, applied in the fields of stem cell biology and cell culture, can solve the problems of unknown serum components, low repeatability, complexity, etc., to improve safety and application value, and reduce batch differences , reducing the effect of differentiation

Inactive Publication Date: 2018-11-16
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 manufacturers, the repeatability of the experiment is extremely low.
In addition, serum may also carry pathogens and other pathogenic factors, which greatly reduces the application value and safety of differentiated nerve cells in drug screening testing or cell therapy.

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  • Serum-free culture system for inducing differentiation of multipotent stem cells into neural progenitor 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] Using DMEM / F12 as the basal medium, add the following ingredients 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 neural progenitor 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 .

[0016] Human induced pluripotent stem cells were cultured in a serum-free minimal medium system to form embryoid bodies. Add 10 μM of SB431542, 1 μM of dermorphin, and 10 μg / L of fibroblast growth factor-2 into the basic medium system, and continu...

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Abstract

The invention belongs to the technical fields of stem cell biology and cell culture, and proposes a serum-free culture system for inducing differentiation of multipotent stem cells into neural progenitor cells. The serum-free culture system concretely comprises a serum-free differentiation base culture medium and an additive promoting differentiation of the neural progenitor cells, wherein the serum-free base culture medium comprises the following essential ingredients: a DMEM/F12 culture medium, human serum albumin, L-glutamine, D-glucose, vitamin C, an N2/B27 cell culture additive and non-essential amino acid; and the additive promoting differentiation of the neural progenitor cells concretely comprises SB431542, dermorphin, fibroblast growth factor-2, retinoic acid and a brain-derived neurotrophic factor. The culture medium provided by the invention can help to increase differentiation of the neural progenitor cells in a cell mixed system and reduce differentiation of the neural progenitor cells into other types of cells, so high-purity neural progenitor cells are finally obtained.

Description

technical field [0001] The invention belongs to the technical fields of stem cell biology and cell culture, and in particular relates to a serum-free culture system for promoting the differentiation of induced pluripotent stem cells to neural progenitor 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, ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797C12N5/071
CPCC12N5/0623C12N2500/32C12N2500/38C12N2500/90C12N2501/115C12N2501/13C12N2501/15C12N2501/85C12N2506/45
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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