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Bioimpacts. 2018;8(2): 129-138.
doi: 10.15171/bi.2018.15
PMID: 29977834
PMCID: PMC6026523
Scopus ID: 85048143913
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Original Research

Reprogramming of mouse fibroblasts into neural lineage cells using biomaterials

Fahsai Kantawong 1*, Chanidapa Saksiriwisitkul 1, Chanakan Riyapa 1, Suchalinee Limpakdee 1, Phenphichar Wanachantararak 2, Thasaneeya Kuboki 3

1 Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand
2 The Dental Research Center, Faculty of Dentistry, Chiang Mai University, Chiang Mai 50200, Thailand
3 Laboratory of Biomedical and Biophysical Chemistry, Institute for Materials Chemistry and Engineering, Kyushu University, Fukuoka, Japan
*Corresponding Author: Email: fahsai.k@cmu.ac.th

Abstract

Introduction: Induced neural stem cells (iNSCs) have the ability of differentiation into neurons, astrocytes and oligodendrocytes. iNSCs are very useful in terms of research and treatment. The present study offers an idea that biomaterials could be one of the tools that could modulate reprogramming process in the fibroblasts.
Methods: Gelatin biomaterials were fabricated into 3 types, including (i) gelatin, (ii) gelatin with 1 mg/mL hydroxyapatite, and (iii) gelatin with hydroxyapatite and pig brain. NIH/3T3 fibroblasts were cultured on each type of biomaterial for 7, 9 and 14 days. RT-PCR was performed to investigate the gene expression of the fibroblasts on biomaterials compared to the fibroblasts on tissue culture plates. PI3K/Akt signaling was performed by flow cytometry after 24 hours seeding on the biomaterials. The biomaterials were also tested with the human APCs and PDL cells.

Results: The fibroblasts exhibited changes in the expression of the reprogramming factor; Klf‫4 and the neural transcription factors; NFIa, NFIb and Ptbp1 after 9 days culture. The cultivation of fibroblasts on the biomaterials for 7 days showed a higher expression of the transcription factor SOX9. The expression of epigenetic genes; Kat2a and HDAC3 were changed upon the cultivation on the biomaterials for 9 days. The fibroblasts cultured on the biomaterials showed an activation of PI3K/Akt signaling. The human APCs and human PDL cells developed mineralization process on biomaterials
Conclusion: Changes in the expression of Klf4, NFIa, NFIb, Ptbp1 and SOX9 indicated that fibroblasts were differentiated into an astrocytic lineage. It is possible that the well-designed biomaterials could work as powerful tools in the reprogramming process of fibroblasts into iNSCs.


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Submitted: 04 Aug 2017
Revision: 30 Dec 2017
Accepted: 04 Jan 2018
ePublished: 10 Jan 2018
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