Mitochondria Regulate the Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth

Abstract

ABSTRACT. Stem cells from human exfoliated deciduous teeth (SHED) are isolated from the dental pulp tissue of primary teeth and can differentiate into neuronal cells. Although SHED are a desirable type of stem cells for transplantation therapy and for the study of neurological diseases, a large part of the neuronal differentiation machinery of SHED remains unclear. Recent studies have suggested that mitochondrial activity is involved in the differentiation of stem cells. In the present work, we investigated the neuronal differentiation machinery of SHED by focusing on mitochondrial activity. During neuronal differentiation of SHED, we observed increased mitochondrial membrane potential, increased mitochondrial DNA, and elongated mitochondria. Furthermore, to examine the demand for mitochondrial activity in neuronal differentiation, we then differentiated SHED into neuronal cells in the presence of rotenone, an inhibitor of mitochondrial respiratory chain complex I, and carbonyl cyanide m-chlorophenyl hydrazone (CCCP), a mitochondrial uncoupler, and found that neuronal differentiation was inhibited by treatment with rotenone and CCCP. These results indicated that increased mitochondrial activity was crucial for the neuronal differentiation of SHED.

Role of pyruvate kinase M2-mediated metabolic reprogramming during podocyte differentiation

“…Cell differentiation was often accompanied by mitochondrial remodeling 28,29 . In order to investigate whether mitochondrial metabolism was associated with podocyte differentiation, mitochondrial morphology was first examined.…”

Accelerated dentinogenesis by inhibiting the mitochondrial fission factor, dynamin related protein 1

“…Mitochondrial activity increases during the differentiation of various stem cells [14,15]. We showed previously that dihydoorotate dehydrogenase, which interacts with the mitochondrial respiratory chain, is important for the differentiation of an osteoblast precursor cell line into osteoblasts [16].…”

Impaired neurite development associated with mitochondrial dysfunction in dopaminergic neurons differentiated from exfoliated deciduous tooth-derived pulp stem cells of children with autism spectrum disorder

“…None of these 3 ASD children showed comorbidities with chromosomal defects, epilepsy or syndromic phenotypes of tuberous sclerosis, Rett syndrome and other Mendelian disorders. The SHED were isolated as previously described [16] , and were cultured in Alpha Modification of Eagle's Medium (Sigma-Aldrich, MO, USA) containing 15% fetal bovine serum (Sigma-Aldrich), 100 µM L -ascorbic acid 2-phosphate (Wako Pure Chemical Industries, Osaka, Japan), 2 mM L -glutamine (Life Technologies, NY, USA), 250 µg/mL fungizone (Life Technologies), 100 U/mL penicillin, and 100 µg/mL streptomycin (Life Technologies), at 37 °C, in an atmosphere containing 5% CO 2 . The cells were used for further experiments as a heterogeneous cell population, according to the experimental procedures reported by previous studies [9] , [10] , [11] .…”

“…Mitochondrial membrane potential (MMP) was measured by staining with JC-1 (Life Technologies), as previously described [16] . Populations of 10,000 cells were analyzed for each sample, and green and red JC-1 signals were detected using the FL1 and FL2 channels, respectively, of a FACSCalibur (BD Bioscience, CA, USA) flow cytometer.…”

“…The expression of transcription factors that are involved in the specification of the midbrain dopaminergic lineage, pituitary homeobox 3 (PITX3) [18] , [19] , and nuclear receptor related 1 protein (NURR1) [20] , [21] were measured using RT-qPCR. Total RNA extraction and RT-qPCR were performed as previously described [16] . The sequences of the primer sets used in this study were as follows: PITX3 , 5′-CCTACGAGGAGGTGTACCCC-3′ (forward) and 5′-AGGCGAATGGAAAGGTCTTGG-3′ (reverse); NURR1 , 5′-GCACTTCGGCAGAGTTGAATGA-3′ (forward) and 5′-GGTGGCTGTGTTGCTGGTAGTT-3′ (reverse); and GAPDH , 5′-GCACCGTCAAGGCTGAGAAC-3′ (forward) and 5′-ATGGTGGTGAAGACGCCAGT-3′ (reverse).…”