Activity of p53 in human amniotic fluid stem cells increases their potentiality as a candidate for stem cell therapy

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.

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

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.623886 ◽

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.

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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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175 HUMAN AMNIOTIC FLUID STEM CELL THERAPY FOR URETHRAL SPHINCTER REGENERATION

The Journal of Urology ◽

10.1016/j.juro.2011.02.244 ◽

2011 ◽

Vol 185 (4S) ◽

Author(s):

Tae Gyun Kwon ◽

Bum Soo Kim ◽

Seock Hwan Choi ◽

Tae Hwan Kim ◽

Eun Sang Yoo ◽

...

Keyword(s):

Stem Cell ◽

Cell Therapy ◽

Amniotic Fluid ◽

Stem Cell Therapy ◽

Urethral Sphincter ◽

Human Amniotic Fluid ◽

Amniotic Fluid Stem Cell

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Mesenchymal stem cells: Stem cell therapy perspectives for type 1 diabetes

Diabetes & Metabolism ◽

10.1016/j.diabet.2008.10.003 ◽

2009 ◽

Vol 35 (2) ◽

pp. 85-93 ◽

Cited By ~ 89

Author(s):

L. Vija ◽

D. Farge ◽

J.-F. Gautier ◽

P. Vexiau ◽

C. Dumitrache ◽

...

Keyword(s):

Stem Cells ◽

Type 1 Diabetes ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Cell Therapy ◽

Stem Cell Therapy

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The promises of stem cells: stem cell therapy for movement disorders

Parkinsonism & Related Disorders ◽

10.1016/s1353-8020(13)70031-2 ◽

2014 ◽

Vol 20 ◽

pp. S128-S131 ◽

Cited By ~ 11

Author(s):

Hideki Mochizuki ◽

Chi-Jing Choong ◽

Toru Yasuda

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Movement Disorders

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Human Amniotic Fluid Mesenchymal Stem Cells from Second- and Third-Trimester Amniocentesis: Differentiation Potential, Molecular Signature, and Proteome Analysis

Stem Cells International ◽

10.1155/2015/319238 ◽

2015 ◽

Vol 2015 ◽

pp. 1-15 ◽

Cited By ~ 36

Author(s):

Jurate Savickiene ◽

Grazina Treigyte ◽

Sandra Baronaite ◽

Giedre Valiuliene ◽

Algirdas Kaupinis ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Amniotic Fluid ◽

Expression Patterns ◽

Specific Gene ◽

Specific Cell ◽

Third Trimester ◽

Human Amniotic Fluid ◽

Differentiation Potential

Human amniotic fluid stem cells have become an attractive stem cell source for potential applications in regenerative medicine and tissue engineering. The aim of this study was to characterize amniotic fluid-derived mesenchymal stem cells (AF-MSCs) from second- and third-trimester of gestation. Using two-stage protocol, MSCs were successfully cultured and exhibited typical stem cell morphological, specific cell surface, and pluripotency markers characteristics. AF-MSCs differentiated into adipocytes, osteocytes, chondrocytes, myocytes, and neuronal cells, as determined by morphological changes, cell staining, and RT-qPCR showing the tissue-specific gene presence for differentiated cell lineages. Using SYNAPT G2 High Definition Mass Spectrometry technique approach, we performed for the first time the comparative proteomic analysis between undifferentiated AF-MSCs from late trimester of gestation and differentiated into myogenic, adipogenic, osteogenic, and neurogenic lineages. The analysis of the functional and expression patterns of 250 high abundance proteins selected from more than 1400 demonstrated the similar proteome of cultured and differentiated AF-MSCs but the unique changes in their expression profile during cell differentiation that may help the identification of key markers in differentiated cells. Our results provide evidence that human amniotic fluid of second- and third-trimester contains stem cells with multilineage potential and may be attractive source for clinical applications.

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Optimizing Stem Cell Therapy after Ischemic Brain Injury

Journal of Stroke ◽

10.5853/jos.2019.03048 ◽

2020 ◽

Vol 22 (3) ◽

pp. 286-305 ◽

Cited By ~ 1

Author(s):

Shuai Zhang ◽

Brittany Bolduc Lachance ◽

Bilal Moiz ◽

Xiaofeng Jia

Keyword(s):

Stem Cells ◽

Clinical Trials ◽

Cardiac Arrest ◽

Stem Cell ◽

Brain Injury ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Ischemic Brain Injury ◽

Ischemic Brain ◽

Injury Treatment

Stem cells have been used for regenerative and therapeutic purposes in a variety of diseases. In ischemic brain injury, preclinical studies have been promising, but have failed to translate results to clinical trials. We aimed to explore the application of stem cells after ischemic brain injury by focusing on topics such as delivery routes, regeneration efficacy, adverse effects, and in vivo potential optimization. PUBMED and Web of Science were searched for the latest studies examining stem cell therapy applications in ischemic brain injury, particularly after stroke or cardiac arrest, with a focus on studies addressing delivery optimization, stem cell type comparison, or translational aspects. Other studies providing further understanding or potential contributions to ischemic brain injury treatment were also included. Multiple stem cell types have been investigated in ischemic brain injury treatment, with a strong literature base in the treatment of stroke. Studies have suggested that stem cell administration after ischemic brain injury exerts paracrine effects via growth factor release, blood-brain barrier integrity protection, and allows for exosome release for ischemic injury mitigation. To date, limited studies have investigated these therapeutic mechanisms in the setting of cardiac arrest or therapeutic hypothermia. Several delivery modalities are available, each with limitations regarding invasiveness and safety outcomes. Intranasal delivery presents a potentially improved mechanism, and hypoxic conditioning offers a potential stem cell therapy optimization strategy for ischemic brain injury. The use of stem cells to treat ischemic brain injury in clinical trials is in its early phase; however, increasing preclinical evidence suggests that stem cells can contribute to the down-regulation of inflammatory phenotypes and regeneration following injury. The safety and the tolerability profile of stem cells have been confirmed, and their potent therapeutic effects make them powerful therapeutic agents for ischemic brain injury patients.

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Stem cells in the treatment of patients with coronary heart disease. Part I

CARDIOVASCULAR THERAPY AND PREVENTION ◽

10.15829/1728-8800-2011-2-122-128 ◽

2011 ◽

Vol 10 (2) ◽

pp. 122-128 ◽

Cited By ~ 1

Author(s):

N. S. Zhukova ◽

I. I. Staroverov

Keyword(s):

Stem Cells ◽

Coronary Heart Disease ◽

Stem Cell ◽

Heart Disease ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Myocardial Damage ◽

Cellular Cardiomyoplasty ◽

Modern Methods ◽

Death Causes

Heart failure (HF) is one of the leading death causes in patients with myocardial infarction (MI). The modern methods of reperfusion MI therapy, such as thrombolysis, surgery and balloon revascularization, even when performed early, could fail to prevent the development of large myocardial damage zones, followed by HF. Therefore, the researches have been searching for the methods which improve functional status of damaged myocardium. This review is focused on stem cell therapy, a method aimed at cardiac function restoration. The results of experimental and clinical studies on stem cell therapy in coronary heart disease are presented. Various types of stem cells, used for cellular cardiomyoplasty, are characterised. The methods of cell transplantation into myocardium and potential adverse effects of stem cell therapy are discussed.

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