scholarly journals Metabolic Profile and Neurogenic Potential of Human Amniotic Fluid Stem Cells From Normal vs. Fetus-Affected Gestations

2021 ◽  
Vol 9 ◽  
Author(s):  
Giedrė Valiulienė ◽  
Aistė Zentelytė ◽  
Elizabet Beržanskytė ◽  
Rūta Navakauskienė
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
Embryonic Stem ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Amniotic Fluid ◽  
Metabolic Potential ◽  
Differentiation Potential ◽  
Amniotic Fluid Stem Cells ◽  
Inflammatory Status

Human amniotic fluid stem cells (hAFSCs) possess some characteristics with mesenchymal stem cells (MSCs) and embryonic stem cells and have a broader differentiation potential compared to MSCs derived from other sources. Although hAFSCs are widely researched, their analysis mainly involves stem cells (SCs) obtained from normal, fetus-unaffected gestations. However, in clinical settings, knowledge about hAFSCs from normal gestations could be poorly translational, as hAFSCs from healthy and fetus-diseased gestations may differ in their differentiation and metabolic potential. Therefore, a more thorough investigation of hAFSCs derived from pathological gestations would provide researchers with the knowledge about the general characteristics of these cells that could be valuable for further scientific investigations and possible future clinical applicability. The goal of this study was to look into the neurogenic and metabolic potential of hAFSCs derived from diseased fetuses, when gestations were concomitant with polyhydramnios and compare them to hAFSCs derived from normal fetuses. Results demonstrated that these cells are similar in gene expression levels of stemness markers (SOX2, NANOG, LIN28A, etc.). However, they differ in expression of CD13, CD73, CD90, and CD105, as flow cytometry analysis revealed higher expression in hAFSCs from unaffected gestations. Furthermore, hAFSCs from “Normal” and “Pathology” groups were different in oxidative phosphorylation rate, as well as level of ATP and reactive oxygen species production. Although the secretion of neurotrophic factors BDNF and VEGF was of comparable degree, as evaluated with enzyme-linked immunosorbent assay (ELISA) test, hAFSCs from normal gestations were found to be more prone to neurogenic differentiation, compared to hAFSCs from polyhydramnios. Furthermore, hAFSCs from polyhydramnios were distinguished by higher secretion of pro-inflammatory cytokine TNFα, which was significantly downregulated in differentiated cells. Overall, these observations show that hAFSCs from pathological gestations with polyhydramnios differ in metabolic and inflammatory status and also possess lower neurogenic potential compared to hAFSCs from normal gestations. Therefore, further in vitro and in vivo studies are necessary to dissect the potential of hAFSCs from polyhydramnios in stem cell-based therapies. Future studies should also search for strategies that could improve the characteristics of hAFSCs derived from diseased fetuses in order for those cells to be successfully applied for regenerative medicine purposes.

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

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
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.

291 ISOLATION AND CHARACTERIZATION OF MESENCHYMAL STEM CELLS DERIVED FROM HUMAN AMNIOTIC FLUID

2011 ◽  
Vol 23 (1) ◽  
pp. 243 ◽  
Author(s):  
S.-A. Choi ◽  
J.-H. Lee ◽  
K.-J. Kim ◽  
E.-Y. Kim ◽  
K.-S. Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Amniotic Fluid ◽  
Adult Stem Cells ◽  
Embryonic Stem ◽  
Stem Cell Marker ◽  
Second Trimester ◽  
Cell Marker ◽  
Human Amniotic Fluid ◽  
Amniotic Fluid Stem Cells

Adult stem cells have the capacity to differentiate into several different cell types, although their differentiation potential is limited compared with that of embryonic stem cells. Thus, adult stem cells are regarded as an exciting source for new cell therapies. Recent observations also indicate that stem cells derived from second-trimester amniocentesis are pluripotent – capable of differentiating into multiple lineages, including representatives of all 3 embryonic germ layers. In addition, amniotic fluid stem cells can be used in the generation of disease- or patient-specific stem cells, and amniotic fluid stem cells could be an ideal source for autologous cell replacement therapy in the later life of the fetus. The aim of the present study was to investigate isolation and characterisation of human amniotic fluid-derived mesenchymal stem cells (hAFS). We successfully isolated and characterised hAFS. Amniotic fluid samples were collected in the second trimester (median gestational age: 16 weeks, range: 15–17 weeks) for prenatal diagnosis. Specimens (2 mL) were centrifuged and incubated in low-glucose DMEM supplemented with 10% FBS, 25 ng of basic fibroblast growth factor, and 10 ng of epidermal growth factor at 37°C with 5% CO2. Human amniotic fluid cell (passage 6) expression of stem cell specific markers OCT-4, SOX2, Rex1, FGF4, and NANOG was confirmed by RT-PCR. Flow cytometric analysis showed that hAFS (passage 10) were positive for CD44, CD29, CD146, STRO1, and CD90 but negative for CD19. Immunocytochemical analysis of hAFS (passage 11) also showed the expression of OCT-4, SSEA-1, CD44, CD29, CD146, STRO1, and CD90, but hAFS were negative for CD19 and CD14. In conclusion, according to the previous studies on other mammalians, hAFS are an appropriate source of pluripotent stem cells. Here, we demonstrated that hAFS have a high expression of stem cell specific marker, including embryonic stem cell marker and mesenchymal stem cell marker. Therefore, amniotic fluid may be a suitable alternative source of multipotent stem cells.

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

2009 ◽  
Vol 18 (7) ◽  
pp. 1003-1012 ◽  
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

scholarly journals Small Molecule Treatments Improve Differentiation Potential of Human Amniotic Fluid Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Aistė Zentelytė ◽  
Deimantė Žukauskaitė ◽  
Ieva Jacerytė ◽  
Veronika V. Borutinskaitė ◽  
Rūta Navakauskienė
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Stem Cell ◽  
Amniotic Fluid ◽  
Small Molecules ◽  
Small Molecule ◽  
Human Amniotic Fluid ◽  
Short Term ◽  
Differentiation Potential ◽  
Amniotic Fluid Stem Cells

Human amniotic fluid stem cells (AFSC) are an exciting and very promising source of stem cells for therapeutic applications. In this study we investigated the effects of short-term treatments of small molecules to improve stem cell properties and differentiation capability. For this purpose, we used epigenetically active compounds, such as histone deacetylase inhibitors Trichostatin A (TSA) and sodium butyrate (NaBut), as well as multifunctional molecules of natural origin, such as retinoic acid (RA) and vitamin C (vitC). We observed that combinations of these compounds triggered upregulation of genes involved in pluripotency (KLF4, OCT4, NOTCH1, SOX2, NANOG, LIN28a, CMYC), but expression changes of these proteins were mild with only significant downregulation of Notch1. Also, some alterations in cell surface marker expression was established by flow cytometry with the most explicit changes in the expression of CD105 and CD117. Analysis of cellular energetics performed using Seahorse analyzer and assessment of gene expression related to cell metabolism and respiration (NRF1, HIF1α, PPARGC1A, ERRα, PKM, PDK1, LDHA, NFKB1, NFKB2, RELA, RELB, REL) revealed that small molecule treatments stimulate AFSCs toward a more energetically active phenotype. To induce cells to differentiate toward neurogenic lineage several different protocols including commercial supplements N2 and B27 together with RA were used and compared to the same differentiation protocols with the addition of a pre-induction step consisting of a combination of small molecules (vitC, TSA and RA). During differentiation the expression of several neural marker genes was analyzed (Nestin, MAP2, TUBB3, ALDH1L1, GFAP, CACNA1D, KCNJ12, KCNJ2, KCNH2) and the beneficial effect of small molecule treatment on differentiation potential was observed with upregulated gene expression. Differentiation was also confirmed by staining TUBB3, NCAM1, and Vimentin and assessed by secretion of BDNF. The results of this study provide valuable insights for the potential use of short-term small molecule treatments to improve stem cell characteristics and boost differentiation potential of AFSCs.

scholarly journals In vitro cardiomyogenic potential of human amniotic fluid stem cells

2011 ◽  
Vol 5 (3) ◽  
pp. 220-228 ◽  
Author(s):  
Xuan Guan ◽  
Dawn M. Delo ◽  
Anthony Atala ◽  
Shay Soker
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
Human Amniotic Fluid ◽  
Amniotic Fluid Stem Cells

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

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.

Human amniotic fluid stem cells: neural differentiation in vitro and in vivo

2014 ◽  
Vol 357 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Tullia Maraldi ◽  
Laura Bertoni ◽  
Massimo Riccio ◽  
Manuela Zavatti ◽  
Gianluca Carnevale ◽  
...  
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
Neural Differentiation ◽  
Human Amniotic Fluid ◽  
Amniotic Fluid Stem Cells

Cellular Cardiomyoplasty with Human Amniotic Fluid Stem Cells: In Vitro and In Vivo Studies

2010 ◽  
Vol 16 (6) ◽  
pp. 1925-1936 ◽  
Author(s):  
Yi-Chun Yeh ◽  
Hao-Ji Wei ◽  
Wen-Yu Lee ◽  
Chu-Leng Yu ◽  
Yen Chang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Amniotic Fluid ◽  
In Vivo Studies ◽  
Human Amniotic Fluid ◽  
Cellular Cardiomyoplasty ◽  
Amniotic Fluid Stem Cells

Mini Review; Differentiation of Human Pluripotent Stem Cells into Oocytes

2020 ◽  
Vol 15 (4) ◽  
pp. 301-307 ◽  
Author(s):  
Gaifang Wang ◽  
Maryam Farzaneh
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryonic Stem ◽  
Human Pluripotent Stem Cells ◽  
Human Oocyte ◽  
Differentiation Potential ◽  
Cell Lineages ◽  
Cell Based Therapy ◽  
Induced Pluripotent

Primary Ovarian Insufficiency (POI) is one of the main diseases causing female infertility that occurs in about 1% of women between 30-40 years of age. There are few effective methods for the treatment of women with POI. In the past few years, stem cell-based therapy as one of the most highly investigated new therapies has emerged as a promising strategy for the treatment of POI. Human pluripotent stem cells (hPSCs) can self-renew indefinitely and differentiate into any type of cell. Human Embryonic Stem Cells (hESCs) as a type of pluripotent stem cells are the most powerful candidate for the treatment of POI. Human-induced Pluripotent Stem Cells (hiPSCs) are derived from adult somatic cells by the treatment with exogenous defined factors to create an embryonic-like pluripotent state. Both hiPSCs and hESCs can proliferate and give rise to ectodermal, mesodermal, endodermal, and germ cell lineages. After ovarian stimulation, the number of available oocytes is limited and the yield of total oocytes with high quality is low. Therefore, a robust and reproducible in-vitro culture system that supports the differentiation of human oocytes from PSCs is necessary. Very few studies have focused on the derivation of oocyte-like cells from hiPSCs and the details of hPSCs differentiation into oocytes have not been fully investigated. Therefore, in this review, we focus on the differentiation potential of hPSCs into human oocyte-like cells.

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