Reprogramming of mouse fibroblasts into neural lineage cells using biomaterials

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.

The Potential of Fibroblast Transdifferentiation to Neuron Using Hydrogels

“…The cells might be passed through the neural stem cell (neural progenitor) state because the NIH/3T3 cells expressed the Klf4, a pluripotency marker which is significantly upregulated on the gelatin with hydroxyapatite and pig brain extract during the first week of the cell culture. The PI3K/Akt activation is involved in the early steps of the reprogramming process as it is increased in NIH/3T3-cell-cultured gelatin with hydroxyapatite and pig brain extract [89]. This evidence strongly supports that hydrogels can modulate transcription factors to cause reprogramming.…”

Peak density of immature nerve cells occurs with high‐grade dysplasia in intraductal papillary mucinous neoplasms of the pancreas

“…Amit et al [20] used a different mouse model of head and neck cancer to show that axons infiltrating a tumor from adjacent sensory nerves may be reprogrammed by tumor exosomederived miRNAs and acquire a sympathetic phenotype. Transdifferentiation of fibroblasts into neural precursors has been demonstrated in vitro [46,47], and some in vivo models have succeeded in transdifferentiating supporting astrocytes into functional neurons in mice [48]. Lastly, Mauffrey et al [36] suggested that DCX + neural precursors migrate from the brain into prostate cancers and generate adrenergic neurons.…”

Current progress in bionanomaterials to modulate the epigenome

“…Kantawong et al 44 explored the potential of gelatin-hydroxyapatite-pig brain biomaterials in generating induced neural stem cells (iNSCs) from MEFs. iNSCs are not only capable of differentiating into neurons, but also into glial cells including astrocytes and oligodendrocytes.…”

“…Indeed, culture on the gelatin materials upregulated astrocytic TF ( NFIA , NFIB , SOX9 ) expression even without viral TF infection. 44 Currently, astrocyte generation is slow in vitro , 48 which is a considerable limiting factor for studying neurodegenerative disease. Future studies could focus on further optimising gelatin-based substrates to shorten the differentiation period.…”