Chemical conversion of human lung fibroblasts into neuronal cells

Abstract

It has been previously reported that human embryonic fibroblasts and mouse embryonic fibroblasts can be converted into neuronal cells using chemical agents, along with forced expression specific transcriptional factors. However, the materials required for reprogramming in these approaches presents major technical difficulties and safety concerns. The current study investigated whether a cocktail of small molecules can convert human lung fibroblast cells into neurons. The small molecules valproic acid, CHIR99021, DMH1, Repsox, forskolin, Y-27632 and SP600125 (VCHRFYS) were used to induce MRC-5 cells into neuronal cells in vitro. Neuronal markers were analyzed by immunofluorescence staining. The gene profiles were analyzed by reverse transcription-quantitative polymerase chain reaction. MRC-5 is a human lung fibroblast cell line derived from normal lung tissue of a 14-week-old male fetus. The results of the current study demonstrated that MRC-5 fibroblasts can be directly converted into neuronal cells using a cocktail of seven small molecules (VCHRFYS), with a yield of ~90% Tuj1-positive cells after 7 days of induction. Following a further maturation period, these chemically-induced neurons possessed neuronal morphology and expressed multiple neuron-specific genes. In conclusion, a cocktail of small molecules that can convert fibroblasts MRC-5 cells into functional neurons without the exogenous genetic factors was identified, which has the potential to be useful in neurological disease therapy.

Long Noncoding RNA H19 Contributes to Cholangiocyte Proliferation and Cholestatic Liver Fibrosis in Biliary Atresia

“…Indeed, the correlation analysis showed that hepatic H19 mRNA levels were positively correlated with hepatic TGFB1 mRNA and ACTA2, suggesting that H19 mRNA levels could be used to monitor LF progression in BA patients. Similar to the findings in mouse CLI models, the current study showed that H19 levels were markedly increased in serum exosomes isolated from BA patients compared to those in controls. Given that exosomal and hepatic H19 levels are associated with LF in BA patients, serum exosomal H19 levels may be utilized as a useful marker for monitoring LF progression of BA patients.…”

“…H19 is highly expressed in fetal liver, but repressed significantly after birth. Intriguingly, it has been reported that H19 is up‐regulated in human liver diseases and CLI animal models, including CCl 4 ‐induced liver injury, bile duct ligation (BDL)‐induced cholestatic injury, and the multidrug resistance 2 knockout ( Mdr2 –/– ) mouse model, implicating its important role in disease progression of CLI. Our recent studies indicate that hepatic H19 level is correlated with severity of cholestatic injury in Mdr2 –/– mice .…”

The Roles and Mechanisms of lncRNAs in Liver Fibrosis

“…LncRNA H19 (both hepatic and serum exosomal long noncoding RNA H19) is an imprinted and maternally expressed gene that is conserved between humans and mice and is involved in the regulation of cell proliferation and differentiation [95,96]. A previous study found that H19 is upregulated in CLI animal models and human liver diseases [92,97,98]. H19 plays an important role in the process of liver fibrosis, with inhibition and promotion mediated by different targets.…”

“…As is a master profibrogenic cytokine, TGF-β controls liver physiology and pathology during the initial process of liver injury-inflammation-fibrosis [92,96,97]. TGF-β influences the different biological processes of liver fibrogenesis, including the circadian rhythm, epigenetics, reactive oxygen species generation, metabolism, senescence, EMT, and endothelial to mesenchymal transition [97]. To date, it has been found that lncRNAs cooperate with the TGF-β signalling pathway to regulate HSC activation and hepatic fibrosis.…”

Small Molecule Epigenetic Modulators in Pure Chemical Cell Fate Conversion

“…At the same time, Pei's work revealed that human fibroblasts were able to transdifferentiate into neurons by a different chemical cocktail (VPA, CHIR99021, RepSox, Forskolin, SP600125, Gö6983, and Y-27632) [ 40 ]. It was also reported that human lung fibroblasts could be converted into neurons using a similar small molecule combination, including VPA, CHIR99021, DMH1, RepSox, Forskolin, Y-27632, and SP600125 [ 41 ].…”