Increased In Vitro Osteopotential in SHED Associated with Higher IGF2 Expression When Compared with hASCs
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Mesenchymal stem cell (MSC) osteogenic differentiation potential varies according to factors such as tissue source and cell population heterogeneity. Pre-selection of cell subpopulations harboring higher osteopotential is a promising strategy to achieve a thorough translation of MSC-based therapies to the clinic. Here, we searched for novel molecular markers predictive of osteopotential by comparing MSC populations from two sources harboring different osteogenic potentials. We show that MSCs from human deciduous teeth (SHED) have an intrinsically higher osteogenic potential when compared with MSCs from human adipose tissue (hASCs) under the same in vitro controlled induction system. Transcriptome profiling revealed IGF2 to be one of the top upregulated transcripts before and during early in vitro osteogenic differentiation. Further, exogenous IGF2 supplementation enhanced alkaline phosphatase activity and matrix mineralization, and inhibition of IGF2 lessened these parameters in SHED and hASCs, validating IGF2 as an osteogenic factor in these MSCs. Further, we found IGF2 to be biallelically expressed in SHED, but not in hASCs. We observed a 4 % methylation increase in the imprinting control region within the IGF2-H19 locus in SHED, and this is mainly due to 2 specific CpG sites. Thus, we suggest that IGF2 upregulation in SHED is due to loss of imprinting. This study unravels osteogenic properties in SHED, implying IGF2 as a potential biomarker of MSCs with higher osteopotential, and unveils IGF2 loss-of-imprinting in SHED.
KeywordsSHED hASCs In vitro osteopotential IGF2 Imprinting Biallelic expression
We would like to thank Constancia Gotto Urbani for secretarial assistance, Andressa Gois Morales and Simone Gomes Ferreira for technical support. We also thank Erika Yeh, PhD (University of California, San Francisco) and Luciane Portas Capelo, PhD (Federal University of Sao Paulo, Sao Jose dos Campos) for stimulating discussions. This work was supported by grants from the Brazillian Ministry of Health, Fundação de Amparo à Pesquisa do Estado de São Paulo (CEPID/FAPESP), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
Disclosure of Potential Conflict of Interest
No potential conflict of interest was disclosed.
Roberto Dalto Fanganiello: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing
Felipe Augusto Andre Ishiy: collection and assembly of data, data analysis and interpretation, manuscript writing
Gerson Shigeru Kobayashi: data analysis and interpretation, manuscript writing
Lucas Alvizi: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing
Daniele Yumi Sunaga: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing
Maria Rita Passos-Bueno: conception and design, data analysis and interpretation, manuscript writing, financial support, administrative support
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