In vitro differentiation into insulin-producing β-cells of stem cells isolated from human amniotic fluid and dental pulp

AbstractOxysterols play vital roles in the human body, ranging from cell cycle regulation and progression to dopaminergic neurogenesis. While naïve human mesenchymal stem cells (hMSCs) have been explored to have neurogenic effect, there is still a grey area to explore their regenerative potential after in vitro differentiation. Hence, in the current study, we have investigated the neurogenic effect of 22(R)-hydroxycholesterol (22-HC) on hMSCs obtained from bone marrow, adipose tissue and dental pulp. Morphological and morphometric analysis revealed physical differentiation of stem cells into neuronal cells. Detailed characterization of differentiated cells affirmed generation of neuronal cells in culture. The percentage of generation of non-DA cells in the culture confirmed selective neurogenic potential of 22-HC. We substantiated the efficacy of these cells in neuro-regeneration by transplanting them into Parkinson’s disease Wistar rat model. MSCs from dental pulp had maximal regenerative effect (with 80.20 ± 1.5% in vitro differentiation efficiency) upon transplantation, as shown by various behavioural examinations and immunohistochemical tests. Subsequential analysis revealed that 22-HC yields a higher percentage of functional DA neurons and has differential effect on various tissue-specific primary human MSCs. 22-HC may be used for treating Parkinson’s disease in future with stem cells.

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Nuclear Nox4 Role in Stemness Power of Human Amniotic Fluid Stem Cells

Oxidative Medicine and Cellular Longevity ◽

10.1155/2015/101304 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 8

Author(s):

Tullia Maraldi ◽

Marianna Guida ◽

Manuela Zavatti ◽

Elisa Resca ◽

Laura Bertoni ◽

...

Keyword(s):

Stem Cells ◽

Transcription Factors ◽

Amniotic Fluid ◽

Human Stem Cells ◽

Human Amniotic Fluid ◽

Differentiation Potential ◽

Amniotic Fluid Stem Cells ◽

Target Molecules ◽

Cellular Compartments

Human amniotic fluid stem cells (AFSC) are an attractive source for cell therapy due to their multilineage differentiation potential and accessibility advantages. However the clinical application of human stem cells largely depends on their capacity to expandin vitro, since there is an extensive donor-to-donor heterogeneity. Reactive oxygen species (ROS) and cellular oxidative stress are involved in many physiological and pathophysiological processes of stem cells, including pluripotency, proliferation, differentiation, and stress resistance. The mode of action of ROS is also dependent on the localization of their target molecules. Thus, the modifications induced by ROS can be separated depending on the cellular compartments they affect. NAD(P)H oxidase family, particularly Nox4, has been known to produce ROS in the nucleus. In the present study we show that Nox4 nuclear expression (nNox4) depends on the donor and it correlates with the expression of transcription factors involved in stemness regulation, such as Oct4, SSEA-4, and Sox2. Moreover nNox4 is linked with the nuclear localization of redox sensitive transcription factors, as Nrf2 and NF-κB, and with the differentiation potential. Taken together, these results suggest that nNox4 regulation may have important effects in stem cell capability through modulation of transcription factors and DNA damage.

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Human amniotic fluid-derived and dental pulp-derived stem cells seeded into collagen scaffold repair critical-size bone defects promoting vascularization

Stem Cell Research & Therapy ◽

10.1186/scrt203 ◽

2013 ◽

Vol 4 (3) ◽

pp. 53 ◽

Cited By ~ 48

Author(s):

Tullia Maraldi ◽

Massimo Riccio ◽

Alessandra Pisciotta ◽

Manuela Zavatti ◽

Gianluca Carnevale ◽

...

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Dental Pulp ◽

Critical Size ◽

Collagen Scaffold ◽

Bone Defects ◽

Human Amniotic Fluid

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Human Amniotic Fluid Stem Cells Do Not Differentiate Into Dopamine Neurons In Vitro or After Transplantation In Vivo

Stem Cells and Development ◽

10.1089/scd.2008.0300 ◽

2009 ◽

Vol 18 (7) ◽

pp. 1003-1012 ◽

Cited By ~ 34

Author(s):

Angela E. Donaldson ◽

Jingli Cai ◽

Ming Yang ◽

Lorraine Iacovitti

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Dopamine Neurons ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells

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Osteogenic Differentiation of Human Amniotic Fluid Mesenchymal Stem Cells Is Determined by Epigenetic Changes

Stem Cells International ◽

10.1155/2016/6465307 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 16

Author(s):

Monika Glemžaitė ◽

Rūta Navakauskienė

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Amniotic Fluid ◽

Osteogenic Differentiation ◽

Epigenetic Changes ◽

Human Amniotic Fluid ◽

Polycomb Repressive Complex 2 ◽

Cell Staining

Osteogenic differentiation of human amniotic fluid derived mesenchymal stem cells (AF-MSCs) has been widely studiedin vitroandin vivoas a potential tool for regenerative medicine and tissue engineering. While most of the studies analyze changes in transcriptional profile during differentiation to date there is not much information regarding epigenetic changes in AF-MSCs during differentiation. The aim of our study was to evaluate epigenetic changes during osteogenic differentiation of AF-MS cells. Isolated AF-MSCs were characterized morphologically and osteogenic differentiation was confirmed by cell staining and determining expression of alkaline phosphatase and osteopontin by RT-qPCR. Variation in gene expression levels of pluripotency markers and specific microRNAs were also evaluated. Analysis of epigenetic changes revealed that levels of chromatin modifying enzymes such as Polycomb repressive complex 2 (PRC2) proteins (EZH2 and SUZ12), DNMT1, HDAC1, and HDAC2 were reduced after osteogenic differentiation of AF-MSCs. We demonstrated that the level of specific histone markers keeping active state of chromatin (H3K4me3, H3K9Ac, and others) increased and markers of repressed state of chromatin (H3K27me3) decreased. Our results show that osteogenic differentiation of AF-MSCs is conducted by various epigenetic alterations resulting in global chromatin remodeling and provide insights for further epigenetic investigations in human AF-MSCs.

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Pluripotent stem cells isolated from human amniotic fluid and differentiation into pancreatic β-cells

Journal of Endocrinological Investigation ◽

10.1007/bf03345764 ◽

2009 ◽

Vol 32 (11) ◽

pp. 873-876 ◽

Cited By ~ 14

Author(s):

L. Trovato ◽

R. De Fazio ◽

M. Annunziata ◽

S. Sdei ◽

E. Favaro ◽

...

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Pluripotent Stem Cells ◽

Β Cells ◽

Pancreatic Β Cells ◽

Human Amniotic Fluid

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Isolation of Dental Pulp Stem Cells and their In Vitro Differentiation into Odontoblast-like Cells

Macedonian Journal of Medical Sciences ◽

10.3889/mjms.1857-5773.2011.0142 ◽

2011 ◽

Vol 4 (3) ◽

pp. 253-260 ◽

Cited By ~ 2

Author(s):

Nermeen El-Motaz Bellah Ahmed ◽

Eman Hassan Anwar Aboul-Ezz ◽

Siza Yacoub Zakhary ◽

Tarek Hamed El Badry ◽

Magda Ismail Ramzy

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

In Vitro Differentiation

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In vitro cardiomyogenic potential of human amniotic fluid stem cells

Journal of Tissue Engineering and Regenerative Medicine ◽

10.1002/term.308 ◽

2011 ◽

Vol 5 (3) ◽

pp. 220-228 ◽

Cited By ~ 44

Author(s):

Xuan Guan ◽

Dawn M. Delo ◽

Anthony Atala ◽

Shay Soker

Keyword(s):

Stem Cells ◽

Amniotic Fluid ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells

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Genetic and epigenetic modifications induced by chemotherapeutic drugs: human amniotic fluid stem cells as an in-vitro model

BMC Medical Genomics ◽

10.1186/s12920-019-0595-3 ◽

2019 ◽

Vol 12 (1) ◽

Author(s):

Prabin Upadhyaya ◽

Alessandra Di Serafino ◽

Luca Sorino ◽

Patrizia Ballerini ◽

Marco Marchisio ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Amniotic Fluid ◽

Real Time ◽

In Vitro Model ◽

Chemotherapeutic Agents ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cells ◽

Vitro Model

Abstract Background Bleomycin, etoposide and cisplatin (BEP) are three chemotherapeutic agents widely used individually or in combination with each other or other chemotherapeutic agents in the treatment of various cancers. These chemotherapeutic agents are cytotoxic; hence, along with killing cancerous cells, they also damage stem cell pools in the body, which causes various negative effects on patients. The epigenetic changes due to the individual action of BEP on stem cells are largely unknown. Methods Human amniotic fluid stem cells (hAFSCs) were treated with our in-vitro standardized dosages of BEP individually, for seven days. The cells were harvested after the treatment and extraction of DNA and RNA were performed. Real-time PCR and flow cytometry were conducted for cell markers analysis. The global DNA methylation was quantified using 5mC specific kit and promoter and CpG methylation % through bisulfite conversion and pyrosequencing. Micro- RNAs (miRNAs) were quantified with real-time qPCR. Results The cytotoxic nature of BEP was observed even at low dosages throughout the experiment. We also investigated the change in the expression of various pluripotent and germline markers and found a significant change in the properties of the cells after the treatments. The methylation of DNA at global, promoter and individual CpG levels largely get fluctuated due to the BEP treatment. Several tested miRNAs showed differential expression. No positive correlation between mRNA and protein expression was observed for some markers. Conclusion Cytotoxic chemotherapeutic agents such as BEP were found to alter stem cell properties of hAFSCs. Different methylation profiles change dynamically, which may explain such changes in cellular properties. Data also suggests that the fate of hAFSCs after treatment may depend upon the interplay between the miRNAs. Finally, our results demonstrate that hAFSCs might prove to be a suitable in-vitro model of stem cells to predict genetic and epigenetic modification due to the action of various drugs.

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