In vivo hepatogenic capacity and therapeutic potential of stem cells from human exfoliated deciduous teeth in liver fibrosis in mice

Abstract

IntroductionLiver transplantation is a gold standard treatment for intractable liver diseases. Because of the shortage of donor organs, alternative therapies have been required. Due to their potential to differentiate into a variety of mature cells, stem cells are considered feasible cell sources for liver regeneration. Stem cells from human exfoliated deciduous teeth (SHED) exhibit hepatogenic capability in vitro. In this study, we investigated their in vivo capabilities of homing and hepatocyte differentiation and therapeutic efficacy for liver disorders in carbon tetrachloride (CCl4)-induced liver fibrosis model mice.MethodsWe transplanted SHED into CCl4-induced liver fibrosis model mice through the spleen, and analyzed the in vivo homing and therapeutic effects by optical, biochemical, histological, immunological and molecular biological assays. We then sorted human leukocyte antigen-ABC (HLA-ABC)-positive cells from primary CCl4-damaged recipient livers, and analyzed their fusogenicity and hepatic characteristics by flow cytometric, genomic DNA, hepatocyte-specific gene assays. Furthermore, we examined the treatment effects of HLA-positive cells to a hepatic dysfunction by a secondary transplantation into CCl4-treated mice.ResultsTransplanted SHED homed to recipient livers, and expressed HLA-ABC, human hepatocyte specific antigen hepatocyte paraffin 1 and human albumin. SHED transplantation markedly recovered liver dysfunction and led to anti-fibrotic and anti-inflammatory effects in the recipient livers. SHED-derived HLA-ABC-positive cells that were sorted from the primary recipient liver tissues with CCl4 damage did not fuse with the host mouse liver cells. Sorted HLA-positive cells not only expressed human hepatocyte-specific genes including albumin, cytochrome P450 1A1, fumarylacetoacetase, tyrosine aminotransferase, uridine 5′-diphospho-glucuronosyltransferase, transferrin and transthyretin, but also secreted human albumin, urea and blood urea nitrogen. Furthermore, SHED-derived HLA-ABC-positive cells were secondary transplanted into CCl4-treated mice. The donor cells homed into secondary recipient livers, and expressed hepatocyte paraffin 1 and human albumin, as well as HLA-ABC. The secondary transplantation recovered a liver dysfunction in secondary recipients.ConclusionsThis study indicates that transplanted SHED improve hepatic dysfunction and directly transform into hepatocytes without cell fusion in CCl4-treated mice, suggesting that SHED may provide a feasible cell source for liver regeneration.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0154-6) contains supplementary material, which is available to authorized users.

Pulp stem cells derived from human permanent and deciduous teeth: Biological characteristics and therapeutic applications

“…For purpose of treating acute renal injury induced by intramuscular injection of glycerol, SHED are revealed to incorporate into tubular, intertubular, perivascular, and glomerulus regions of kidney following intravenous or intraperitoneal injection . It was also reported that SHED are capable of differentiating into hepatocyte‐like cells directly without fusion in mouse models of carbon tetrachloride‐induced liver fibrosis, therefore recovering liver disfunction . In a recent mouse in vivo study, dental pulp pluripotent‐like stem cells (DPPSCs), a subset of DPSCs expressing pluripotency markers including OCT4, Sox2, and NANOG, are demonstrated to integrate in αSMA‐positive smooth muscle layer of blood vessels in skin wounds and dystrophic muscles, in concordance with their robust in vitro smooth muscle cell differentiation potential.…”

“…107 118 Additionally, predifferentiated PSCs, including hepatocytes, 125 myocytes, 128 osteoblasts, 143 islet-like cell cluster, 45 and keratocytes, 137 Table 3, such as calvarial defect, 110 cerebral ischemia, 116,117 and limbal stem cell deficiency, 38 It was also reported that SHED are capable of differentiating into hepatocyte-like cells directly without fusion in mouse models of carbon tetrachloride-induced liver fibrosis, therefore recovering liver disfunction. 41 In a recent mouse in vivo study, 12…”

“…Taken together, it is considered that both neural differentiation and paracrine mechanism are responsible for SHED‐induced therapeutic potential for contused spinal cord injury. Likewise, it has been demonstrated that SHED transplantation ameliorate liver fibrotic disfunction via integrated actions of both direct hepatocyte differentiation and indirect paracrine mechanism‐mediated antifibrosis and anti‐inflammation potentials . Consequently, in terms of mechanisms underlying PSC transplantation‐derived benefits, it is important to note that paracrine mechanism initiated by therapeutic factors secreted from transplanted PSCs, at least partially, participates in treatment and enhances endogenous tissue repair capability.…”

Stem cells from exfoliated deciduous teeth alleviate hyposalivation caused by Sjögren syndrome

“…Compared with other human stem cells, SHED represent an interesting stem cell source and are obtained using non-invasive techniques. To date, SHED have been used to treat diseases such as pulp-pulp dentin regeneration (Rosa, Zhang, Grande, & Nor, 2013), jaw regeneration (Jahanbin et al, 2015), nervous system diseases (Taghipour et al, 2012), immune system diseases (Ma et al, 2012), corneal injury (Gomes et al, 2010), liver injury (Yamaza et al, 2015), lung injury (Wakayama et al, 2015), kidney injury (Hattori et al, 2015) and diabetes (Kanafi et al, 2013). However, it is unknown whether SHED have effect on the hyposalivation caused by SS.…”

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

“…Exfoliated deciduous teeth are usually discarded as medical waste; therefore, SHED can be easily collected from exfoliated deciduous teeth with cooperation and agreement from the patient. SHED express embryonic stem cell markers Oct-4 and Nanog, the mesenchymal stem cell marker stromal precursor antigen-1 (STRO-1), and the neuronal stem cell marker Nestin and possess high prolifer-ation capacity; SHED also show multidifferentiation potential and can differentiate into neuronal, osteogenic, adipogenic, and hepatogenic cells (Ma et al, 2012;Miura et al, 2003;Yamaza et al, 2015). SHED have been shown to be able to differentiate into neurons, astrocytes, and oligodendrocytes (Jarmalavičiūtė et al, 2013;Miura et al, 2003;Nourbakhsh et al, 2011;Sakai et al, 2012).…”