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. 2024 Mar 21;25(6):3546.
doi: 10.3390/ijms25063546.

Modeling Lysosomal Storage Disorders in an Innovative Way: Establishment and Characterization of Stem Cell Lines from Human Exfoliated Deciduous Teeth of Mucopolysaccharidosis Type II Patients

Sofia Carvalho  1   2   3   4 , Juliana Inês Santos  1   2   3   5 , Luciana Moreira  1   2   3 , Ana Joana Duarte  1   2   3   6 , Paulo Gaspar  7 , Hugo Rocha  7 , Marisa Encarnação  1   2   3 , Diogo Ribeiro  1   2   3 , Matilde Barbosa Almeida  1   2   3   8 , Mariana Gonçalves  1   2   3   9 , Hugo David  1   2   3   5 , Liliana Matos  1   2   3 , Olga Amaral  1   2   3 , Luísa Diogo  10 , Sara Ferreira  10 , Constança Santos  10 , Esmeralda Martins  11 , Maria João Prata  5   12 , Luís Pereira de Almeida  4 , Sandra Alves  1   2   3 , Maria Francisca Coutinho  1   2   3
Affiliations

Modeling Lysosomal Storage Disorders in an Innovative Way: Establishment and Characterization of Stem Cell Lines from Human Exfoliated Deciduous Teeth of Mucopolysaccharidosis Type II Patients

Sofia Carvalho et al. Int J Mol Sci. .

Abstract

Among the many lysosomal storage disorders (LSDs) that would benefit from the establishment of novel cell models, either patient-derived or genetically engineered, is mucopolysaccharidosis type II (MPS II). Here, we present our results on the establishment and characterization of two MPS II patient-derived stem cell line(s) from deciduous baby teeth. To the best of our knowledge, this is the first time a stem cell population has been isolated from LSD patient samples obtained from the dental pulp. Taking into account our results on the molecular and biochemical characterization of those cells and the fact that they exhibit visible and measurable disease phenotypes, we consider these cells may qualify as a valuable disease model, which may be useful for both pathophysiological assessments and in vitro screenings. Ultimately, we believe that patient-derived dental pulp stem cells (DPSCs), particularly those isolated from human exfoliated deciduous teeth (SHEDs), may represent a feasible alternative to induced pluripotent stem cells (iPSCs) in many labs with standard cell culture conditions and limited (human and economic) resources.

Keywords: dental pulp stem cells (DPSCs); disease modeling; in vitro models; induced pluripotent stem cells (iPSCs); mucopolysaccharidosis type II; stem cells from human exfoliated deciduous teeth (SHEDs).

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Optical microscope images (4×) of an MPS II-derived SHED primary culture. Undifferentiated SHEDs have a spindle shape similar to fibroblasts. (a) Fusiform format; (b) small colony formation (CFU-F); (c) sub-confluence. Cells were observed using a Leica DMIL inverted contrasting microscope (Leica Microsystems, Wetzlar, Germany) with 4× magnification.
Figure 2
Figure 2
Immunostaining of MPS II-derived SHEDs. (a) Confirmation of 3-germ-layer differentiation capacity: mesoderm (top), endoderm (middle), ectoderm (down). (b) Confirmation of neural progenitor cell (NPC) stage of SHEDs: Nestin and Sox-1 (top line); Pax-6 and Sox-2 (bottom line). Scale bar: 50 µm. Images were acquired using a DM400 M fluorescence microscope (Leica, Wetzlar, Germany).
Figure 3
Figure 3
Biochemical profile of SHEDs. (a) IDS enzyme activity. Data are presented as means ± SEM (n = 3) *** p < 0.0005. (b) HS and DS levels of control- and MPS II-derived SHEDs. (c) Representative confocal images of control (left) and MPS II-derived SHEDs (middle and right) immunostained for lysosomal-associated membrane protein 1 (LAMP1). Scale bar: 25 µm. The outlined regions (white squares) highlight the different lysosomal positioning in controls vs. patient-derived SHEDs, with more pronounced LAMP1 staining in the perinuclear region of MPS II SHEDs. Images were acquired using a TCS-SPE confocal microscope (Leica, Wetzlar, Germany).
Figure 4
Figure 4
Schematic drawing illustrating the different sources of dental mesenchymal stem cells (DMSCs) in the oral cavity. Abbreviations: GMSCs, gingiva-derived mesenchymal stem cells; DFSCs, dental follicle stem cells; SHEDs, stem cells from exfoliated deciduous teeth; SCAPs, stem cells from the apical papilla; DPSCs, dental pulp stem cells; PDLSCs, periodontal ligament stem cells (reproduced from [22]).
See this image and copyright information in PMC

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