Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors

Hua Qin; An-Dong Zhao; Meng-Li Sun; Kui Ma; Xiao-Bing Fu

doi:10.1186/s40779-020-00284-2

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Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors

RESEARCH | Updated：2023-02-28

- Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors
- Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors
- 解放军医学杂志（英文版） 2021年8卷第2期页码：162-173
- Affiliations：
  
  1.Research Center for Tissue Repair and Regeneration affiliated to the Medical Innovation Research Division and 4th Medical Center, PLA General Hospital and PLA Medical College, 28 Fu Xing Road, Haidian District, Beijing 100853, China
  2.Tianjin Medical University, Tianjin 300070, China
  3.PLA Key Laboratory of Tissue Repair and Regenerative Medicine and Beijing Key Research Laboratory of Skin Injury, Repair and Regeneration, Beijing 100048, China
  4.Research Unit of Trauma Care, Tissue Repair and Regeneration, Chinese Academy of Medical Sciences, 2019RU051, Beijing 100048, China
- Author bio：
  
  * fuxiaobing@vip.sina.com
- Funds：
  
  the National Nature Science Foundation of China(81830064;81721092);the National Key Research and Development Plan(2017YFC1103304);the CAMS Innovation Fund for Medical Sciences(CIFMS, 2019-I2M-5-059);the Military Medical Research and Development Projects(AWS17J005;2019–126)
- DOI：10.1186/s40779-020-00284-2
  中图分类号：
- 纸质出版：2021-06
- Accepted：
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Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors[J]. 解放军医学杂志（英文版）, 2021,8(2):162-173.

Qin et al.: Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors. Mil Med Res, 2020, 7: 52.
Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors[J]. 解放军医学杂志（英文版）, 2021,8(2):162-173. DOI： 10.1186/s40779-020-00284-2.

Qin et al.: Direct conversion of human fibroblasts into dopaminergic neuron-like cells using small molecules and protein factors. Mil Med Res, 2020, 7: 52. DOI： 10.1186/s40779-020-00284-2.

摘要

Abstract

Background:

Generation of neurons is essential in cell replacement therapy for neurodegenerative disorders like Parkinson’s disease. Several studies have reported the generation of dopaminergic (DA) neurons from mouse and human fibroblasts by ectopic expression of transcription factors

in which genetic manipulation is associated with potential risks.

Methods:

The small molecules and protein factors were selected based on their function to directly induce human fetal lung IMR-90 fibroblasts into DA neuron-like cells. Microscopical

immunocytochemical

and RT-qPCR analyses were used to characterize the morphology

phenotype

and gene expression features of the induced cells. The whole-cell patch-clamp recordings were exploited to measure the electrophysiological properties.

Results:

Human IMR-90 fibroblasts were rapidly converted into DA neuron-like cells after the chemical induction using small molecules and protein factors

with a yield of approximately 95% positive TUJ1-positive cells. The induced DA neuron-like cells were immunopositive for pan-neuronal markers MAP2

NEUN

and Synapsin 1 and DA markers TH

DDC

DAT

and NURR1. The chemical induction process did not involve a neural progenitor/stem cell intermediate stage. The induced neurons could fire single action potentials

which reflected partially the electrophysiological properties of neurons.

Conclusions:

We developed a chemical cocktail of small molecules and protein factors to convert human fibroblasts into DA neuron-like cells without passing through a neural progenitor/stem cell intermediate stage. The induced DA neuron-like cells from human fibroblasts might provide a cellular source for cell-based therapy of Parkinson’s disease in the future.

关键词

Keywords

references

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