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Article

Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations

by
Francesca Casciaro
1,2,3,
Silvia Zia
4,
Mattia Forcato
5,
Manuela Zavatti
1,
Francesca Beretti
1,
Emma Bertucci
6,
Andrea Zattoni
7,
Pierluigi Reschiglian
7,
Francesco Alviano
8,
Laura Bonsi
8,
Matilde Yung Follo
2,
Marco Demaria
3,
Barbara Roda
7,* and
Tullia Maraldi
1
1
Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41124 Modena, Italy
2
Cellular Signalling Laboratory, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy
3
European Research Institute for the Biology of Ageing (ERIBA), University Medical Center Groningen (UMCG), University of Groningen, 9713 Groningen, The Netherlands
4
Stem Sel srl., 40127 Bologna, Italy
5
Department of Life Sciences, University of Modena and Reggio Emilia, 41124 Modena, Italy
6
Department of Medical and Surgical Sciences for Mothers, Children and Adults, University of Modena and Reggio Emilia, Azienda Ospedaliero Universitaria Policlinico, 41124 Modena, Italy
7
Department of Chemistry “G. Ciamician”, University of Bologna, 40125 Bologna, Italy
8
Unit of Histology, Embryology and Applied Biology, Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, 40125 Bologna, Italy
*
Author to whom correspondence should be addressed.
Cells 2021, 10(1), 158; https://doi.org/10.3390/cells10010158
Submission received: 10 December 2020 / Revised: 7 January 2021 / Accepted: 8 January 2021 / Published: 15 January 2021
(This article belongs to the Section Stem Cells)
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Abstract

Human amniotic fluid stem cells (hAFSCs) are broadly multipotent immature progenitor cells with high self-renewal and no tumorigenic properties. These cells, even amplified, present very variable morphology, density, intracellular composition and stemness potential, and this heterogeneity can hinder their characterization and potential use in regenerative medicine. Celector® (Stem Sel ltd.) is a new technology that exploits the Non-Equilibrium Earth Gravity Assisted Field Flow Fractionation principles to characterize and label-free sort stem cells based on their solely physical characteristics without any manipulation. Viable cells are collected and used for further studies or direct applications. In order to understand the intrapopulation heterogeneity, various fractions of hAFSCs were isolated using the Celector® profile and live imaging feature. The gene expression profile of each fraction was analysed using whole-transcriptome sequencing (RNAseq). Gene Set Enrichment Analysis identified significant differential expression in pathways related to Stemness, DNA repair, E2F targets, G2M checkpoint, hypoxia, EM transition, mTORC1 signalling, Unfold Protein Response and p53 signalling. These differences were validated by RT-PCR, immunofluorescence and differentiation assays. Interestingly, the different fractions showed distinct and unique stemness properties. These results suggest the existence of deep intra-population differences that can influence the stemness profile of hAFSCs. This study represents a proof-of-concept of the importance of selecting certain cellular fractions with the highest potential to use in regenerative medicine.
Keywords: sorting; amniotic fluid stem cells; transcriptome; stemness sorting; amniotic fluid stem cells; transcriptome; stemness
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MDPI and ACS Style

Casciaro, F.; Zia, S.; Forcato, M.; Zavatti, M.; Beretti, F.; Bertucci, E.; Zattoni, A.; Reschiglian, P.; Alviano, F.; Bonsi, L.; et al. Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations. Cells 2021, 10, 158. https://doi.org/10.3390/cells10010158

AMA Style

Casciaro F, Zia S, Forcato M, Zavatti M, Beretti F, Bertucci E, Zattoni A, Reschiglian P, Alviano F, Bonsi L, et al. Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations. Cells. 2021; 10(1):158. https://doi.org/10.3390/cells10010158

Chicago/Turabian Style

Casciaro, Francesca, Silvia Zia, Mattia Forcato, Manuela Zavatti, Francesca Beretti, Emma Bertucci, Andrea Zattoni, Pierluigi Reschiglian, Francesco Alviano, Laura Bonsi, and et al. 2021. "Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations" Cells 10, no. 1: 158. https://doi.org/10.3390/cells10010158

APA Style

Casciaro, F., Zia, S., Forcato, M., Zavatti, M., Beretti, F., Bertucci, E., Zattoni, A., Reschiglian, P., Alviano, F., Bonsi, L., Follo, M. Y., Demaria, M., Roda, B., & Maraldi, T. (2021). Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations. Cells, 10(1), 158. https://doi.org/10.3390/cells10010158

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