Induction of Tyrosine Hydroxylase Gene Expression in Embryonal Carcinoma Stem Cells Using a Natural Tissue‐Specific Inducer

Abstract

A large number of studies have focused on the generation of dopaminergic neurons from pluripotent cells. Differentiation of stem cells into distinct cell types is influenced by tissue-specific microenvironment. Since, central nervous system undergoes further development during postnatal life, in the present study neonatal rat brain tissue extract (NRBE) was applied to direct the differentiation of embryonal carcinoma stem cell line, P19 into dopaminergic (DA) phenotypes. Additionally, a neuroprotective drug, deprenyl was used alone or in combination with the extract. Results from morphological, immunofluorescence, and qPCR analyses showed that during a period of one to three weeks, a large percentage of stem cells were differentiated into neural cells. The results also indicated the greater effect of NRBE on the differentiation of the cells into tyrosine hydroxylase-expressing cells. MS analysis of NRBE showed the enrichment of gene ontology terms related to cell differentiation and neurogenesis. Network analysis of the studied genes and some DA markers resulted in the suggestion of potential regulatory candidates such as AVP, ACHE, LHFPL5, and DLK1 genes. In conclusion, NRBE as a natural native inducer was apparently able to simulate the brain microenvironment and support neural differentiation of P19 cells.

Synergistic Effects of Combined Nurr1 Overexpression and Natural Inducers on the More Efficient Production of Dopaminergic Neuron-Like Cells From Stem Cells

“…In the previous study, we showed that neonatal whole brain extract could induce DA phenotype with high expression of TH gene in P19 cells. In addition, LC-MS/MS analysis confirmed the presence of factors related to neuronal differentiation in NRBE content (Momendoust et al, 2019 ). The existence of cytokines, growth factors, and some other peptides in the developing and adult rat brain tissue has been proved previously (Pettmann et al, 1986 ; Bartlett et al, 1991 ; Lindholm et al, 1992 ; Lee et al, 1993 ; Mogi et al, 1994 ).…”

“…Sox-2 can control pluripotency and neural differentiation of human pluripotent stem cells (Khoo et al, 2013;Zhang and Cui, 2014). Downregulation of Sox-2 in the differentiated cells derived from stem cells has been wellestablished previously (Ebrahimie et al, 2014;Mehrfarjam et al, 2016;Momendoust et al, 2019). IHowever, it is reported that depending on the environmental input, Sox-2 could function as a pluripotency and, or neural differentiation factor (Zhang et al, 2019).…”

“…The rats were mated overnight, and the day neonates were born was considered as P0. One-week-old neonates sacrificed, their whole brains removed from the skull, and brain extracts prepared as described previously (Momendoust et al, 2019 ). Briefly, the whole brain was homogenized in a protease inhibitor (PMSF, Roche, 10 837 091 001) solution and then centrifuged at 3,000 rpm (10 min) and 12,000 rpm (20 min).…”

“…In order to produce highly specialized cells, it will be beneficial to create an environment similar to natural in vivo conditions. Numerous reports have shown that tissue-specific microenvironment can induce stem cell differentiation into appropriate cell types with the desired characteristics (Rivera et al, 2006 ; Bentz et al, 2010 ; Han et al, 2014 ; Narayanan et al, 2014 ; Momendoust et al, 2019 ). Recently, several studies have indicated that the use of a neonatal rat brain extract (NRBE) alone or in combination with a three-dimensional culture system (Azizi et al, 2018 ) or a neuroprotective drug, deprenyl (Momendoust et al, 2019 ), could effectively induce DA phenotype with the ability to express TH gene in stem cells.…”

Trans-differentiation of mouse mesenchymal stem cells into pancreatic β-like cells by a traditional anti-diabetic medicinal herb Medicago sativa L

“…Recently, the effect of neonatal rat brain extract (NRBE) on neural differentiation of an EC stem cell line, P19, was approved. 30 , 31 Furthermore, the effects of neonate mouse pancreas extract (MPE)/conditioned medium (PCM) on the differentiation of P19 cells into pancreatic β-cells have been demonstrated previously. 29 , 32 MPE was also able to induce trans -differentiation of bone marrow mesenchymal stem cells into IPCs.…”