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Salami H, Mowal SJ, Moukhah R, Hajebrahimi Z, Hosseini SA, Edalat H. Evaluating the Differential Effects of Valproic Acid on Wharton's Jelly Mesenchymal Stem Cells. Adv Pharm Bull 2019; 9:497-504. [PMID: 31592436 PMCID: PMC6773934 DOI: 10.15171/apb.2019.059] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/12/2019] [Revised: 03/31/2019] [Accepted: 04/14/2019] [Indexed: 12/31/2022] Open
Abstract
Purpose: The histone deacetylases (HDAC) inhibitor, valproic acid (VPA), is a common
antiepileptic drug and is attractive for its broad range of therapeutic effects on many diseases. It
has been employed as an inducer of pluripotency in some cultured cells. Conversely, VPA has
also been employed as an inducer of in vitro differentiation in many other cells. Therefore, we
employed WJMSCs as a cellular target to evaluate the differential effects of of VPA on potency
state and differentiation level of Wharton’s Jelly mesenchymal stem cells (WJMSCs) in various
concentrations and different culture mediums.
Methods: The isolated WJMSCs were cultured in DMEM (MSC medium). According to previous
protocols, WJMSCs were treated with 0, 0.5 and 1 mM VPA in MSC or embryonic stem cell (ESC)
medium and 2 mM VPA in neural differentiation medium. Real-time polymerase chain reaction
(PCR) and western blot analysis were performed for evaluating the expression of pluripotency
markers. MTT and caspase assays were also performed on VPA-treated cells.
Results: The expression of pluripotency markers and the viability of the WJMSCs – determined
by MTT assay – were significantly increased after 0.5 mM VPA treatment in ESC medium. A 2
mM VPA treatment in neural differentiation medium significantly diminished the expression of
pluripotency markers and the viability of WJMSCs.
Conclusion: According to our results, both VPA concentration and the medium context can
influence VPA effects on WJMSCs. The differential effects of VPA on WJMSCs can reflect its wide
range of effects in the treatment of various diseases.
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Affiliation(s)
- Homa Salami
- Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
| | - Seyed Javad Mowal
- Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
| | - Rasoul Moukhah
- Quality assurance Department, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran
| | - Zahra Hajebrahimi
- Aerospace Research Institute, Ministry of Science, Research and Technology, Tehran, Iran
| | | | - Houri Edalat
- Human Genetics Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
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CRISPR/Cas9-mediated correction of human genetic disease. SCIENCE CHINA-LIFE SCIENCES 2017; 60:447-457. [PMID: 28534256 DOI: 10.1007/s11427-017-9032-4] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/12/2017] [Accepted: 03/05/2017] [Indexed: 12/21/2022]
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Pipino C, Pandolfi A. Osteogenic differentiation of amniotic fluid mesenchymal stromal cells and their bone regeneration potential. World J Stem Cells 2015; 7:681-690. [PMID: 26029340 PMCID: PMC4444609 DOI: 10.4252/wjsc.v7.i4.681] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/12/2014] [Revised: 01/16/2015] [Accepted: 02/11/2015] [Indexed: 02/06/2023] Open
Abstract
In orthopedics, tissue engineering approach using stem cells is a valid line of treatment for patients with bone defects. In this context, mesenchymal stromal cells of various origins have been extensively studied and continue to be a matter of debate. Although mesenchymal stromal cells from bone marrow are already clinically applied, recent evidence suggests that one may use mesenchymal stromal cells from extra-embryonic tissues, such as amniotic fluid, as an innovative and advantageous resource for bone regeneration. The use of cells from amniotic fluid does not raise ethical problems and provides a sufficient number of cells without invasive procedures. Furthermore, they do not develop into teratomas when transplanted, a consequence observed with pluripotent stem cells. In addition, their multipotent differentiation ability, low immunogenicity, and anti-inflammatory properties make them ideal candidates for bone regenerative medicine. We here present an overview of the features of amniotic fluid mesenchymal stromal cells and their potential in the osteogenic differentiation process. We have examined the papers actually available on this regard, with particular interest in the strategies applied to improve in vitro osteogenesis. Importantly, a detailed understanding of the behavior of amniotic fluid mesenchymal stromal cells and their osteogenic ability is desirable considering a feasible application in bone regenerative medicine.
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Pipino C, Mukherjee S, David AL, Blundell MP, Shaw SW, Sung P, Shangaris P, Waters JJ, Ellershaw D, Cavazzana M, Mostoslavsky G, Pandolfi A, Pierro A, Guillot PV, Thrasher AJ, De Coppi P. Trisomy 21 mid-trimester amniotic fluid induced pluripotent stem cells maintain genetic signatures during reprogramming: implications for disease modeling and cryobanking. Cell Reprogram 2014; 16:331-44. [PMID: 25162836 DOI: 10.1089/cell.2013.0091] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022] Open
Abstract
Trisomy 21 is the most common chromosomal abnormality and is associated primarily with cardiovascular, hematological, and neurological complications. A robust patient-derived cellular model is necessary to investigate the pathophysiology of the syndrome because current animal models are limited and access to tissues from affected individuals is ethically challenging. We aimed to derive induced pluripotent stem cells (iPSCs) from trisomy 21 human mid-trimester amniotic fluid stem cells (AFSCs) and describe their hematopoietic and neurological characteristics. Human AFSCs collected from women undergoing prenatal diagnosis were selected for c-KIT(+) and transduced with a Cre-lox-inducible polycistronic lentiviral vector encoding SOX2, OCT4, KLF-4, and c-MYC (50,000 cells at a multiplicity of infection (MOI) 1-5 for 72 h). The embryonic stem cell (ESC)-like properties of the AFSC-derived iPSCs were established in vitro by embryoid body formation and in vivo by teratoma formation in RAG2(-/-), γ-chain(-/-), C2(-/-) immunodeficient mice. Reprogrammed cells retained their cytogenetic signatures and differentiated into specialized hematopoietic and neural precursors detected by morphological assessment, immunostaining, and RT-PCR. Additionally, the iPSCs expressed all pluripotency markers upon multiple rounds of freeze-thawing. These findings are important in establishing a patient-specific cellular platform of trisomy 21 to study the pathophysiology of the aneuploidy and for future drug discovery.
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Affiliation(s)
- Caterina Pipino
- 1 Surgery Unit, Institute of Child Health, University College London , London, WC1N 1EH, United Kingdom
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Spinelli V, Guillot PV, De Coppi P. Induced pluripotent stem (iPS) cells from human fetal stem cells (hFSCs). Organogenesis 2013; 9:101-10. [PMID: 23823661 DOI: 10.4161/org.25197] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023] Open
Abstract
INTRODUCTION (1) Human embryonic stem (ES) cells are pluripotent but are difficult to be used for therapy because of immunological, oncological and ethical barriers. (2) Pluripotent cells exist in vivo, i.e., germ cells and epiblast cells but cannot be isolated without sacrificing the developing embryo. (3) Reprogramming to pluripotency is possible from adult cells using ectopic expression of OKSM and other integrative and non-integrative techniques. (4) Hurdles to overcome include i.e stability of the phenotype in relation to epigenetic memory. SOURCES OF DATA We reviewed the literature related to reprogramming, pluripotency and fetal stem cells. AREAS OF AGREEMENT (1) Fetal stem cells present some advantageous characteristics compared with their neonatal and postnatal counterparts, with regards to cell size, growth kinetics, and differentiation potential, as well as in vivo tissue repair capacity. (2) Amniotic fluid stem cells are more easily reprogrammed to pluripotency than adult fibroblast. (3) The parental population is heterogeneous and present an intermediate phenotype between ES and adult somatic stem cells, expressing markers of both. AREAS OF CONTROVERSY (1) It is unclear whether induced pluripotent stem (iPS) derived from amniotic fluid stem cells are fully or partially reprogrammed. (2) Optimal protocols to ensure highest efficiency and phenotype stability remains to be determined. (3) The "level" of reprogramming, fully vs partial, of iPS derived from amniotic fluid stem cells remain to be determined. GROWING POINTS Banking of fully reprogrammed cells may be important both for (1) autologous and allogenic applications in medicine, and (2) disease modeling.
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Affiliation(s)
- Valentina Spinelli
- Surgery Unit, Institute of Child Health, University College London and Great Ormond Street Hospital, London, UK.
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Moschidou D, Mukherjee S, Blundell MP, Drews K, Jones GN, Abdulrazzak H, Nowakowska B, Phoolchund A, Lay K, Ramasamy TS, Cananzi M, Nettersheim D, Sullivan M, Frost J, Moore G, Vermeesch JR, Fisk NM, Thrasher AJ, Atala A, Adjaye J, Schorle H, De Coppi P, Guillot PV. Valproic acid confers functional pluripotency to human amniotic fluid stem cells in a transgene-free approach. Mol Ther 2012; 20:1953-67. [PMID: 22760542 PMCID: PMC3464631 DOI: 10.1038/mt.2012.117] [Citation(s) in RCA: 110] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/28/2012] [Accepted: 05/21/2012] [Indexed: 12/20/2022] Open
Abstract
Induced pluripotent stem cells (iPSCs) with potential for therapeutic applications can be derived from somatic cells via ectopic expression of a set of limited and defined transcription factors. However, due to risks of random integration of the reprogramming transgenes into the host genome, the low efficiency of the process, and the potential risk of virally induced tumorigenicity, alternative methods have been developed to generate pluripotent cells using nonintegrating systems, albeit with limited success. Here, we show that c-KIT+ human first-trimester amniotic fluid stem cells (AFSCs) can be fully reprogrammed to pluripotency without ectopic factors, by culture on Matrigel in human embryonic stem cell (hESC) medium supplemented with the histone deacetylase inhibitor (HDACi) valproic acid (VPA). The cells share 82% transcriptome identity with hESCs and are capable of forming embryoid bodies (EBs) in vitro and teratomas in vivo. After long-term expansion, they maintain genetic stability, protein level expression of key pluripotency factors, high cell-division kinetics, telomerase activity, repression of X-inactivation, and capacity to differentiate into lineages of the three germ layers, such as definitive endoderm, hepatocytes, bone, fat, cartilage, neurons, and oligodendrocytes. We conclude that AFSC can be utilized for cell banking of patient-specific pluripotent cells for potential applications in allogeneic cellular replacement therapies, pharmaceutical screening, and disease modeling.
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Affiliation(s)
| | | | | | - Katharina Drews
- Max Planck Institute for Molecular Genetics, Berlin, Germany
| | | | | | - Beata Nowakowska
- Center for Human Genetics, Katholieke Universiteit Leuven, Leuven, Belgium
| | | | | | | | | | - Daniel Nettersheim
- Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Bonn, Germany
| | | | | | | | - Joris R Vermeesch
- Center for Human Genetics, Katholieke Universiteit Leuven, Leuven, Belgium
| | - Nicholas M Fisk
- UQ Centre for Clinical Research, University of Queensland, Brisbane, Australia
| | | | - Anthony Atala
- Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, North Carolina, USA
| | - James Adjaye
- Max Planck Institute for Molecular Genetics, Berlin, Germany
- Institute for Stem Cell Research and Regenerative Medicine, Heinrich Heine University Duesseldorf, Duesseldorf, Germany
| | - Hubert Schorle
- Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Bonn, Germany
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Cananzi M, De Coppi P. CD117(+) amniotic fluid stem cells: state of the art and future perspectives. Organogenesis 2012; 8:77-88. [PMID: 23037870 DOI: 10.4161/org.22426] [Citation(s) in RCA: 68] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
Broadly multipotent stem cells can be isolated from amniotic fluid by selection for the expression of the membrane stem cell factor receptor c-Kit, a common marker for multipotential stem cells. They have clonogenic capability and can be directed into a wide range of cell types representing the three primary embryonic lineages. Amniotic fluid stem cells maintained for over 250 population doublings retained long telomeres and a normal karyotype. Clonal human lines verified by retroviral marking were induced to differentiate into cell types representing each embryonic germ layer, including cells of adipogenic, osteogenic, myogenic, endothelial, neuronal and hepatic lineages. AFS cells could be differentiate toward cardiomyogenic lineages, when co-cultured with neonatal cardiomyocytes, and have the potential to generate myogenic and hematopoietic lineages both in vitro and in vivo. Very recently first trimester AFS cells could be reprogrammed without any genetic manipulation opening new possibilities in the field of fetal/neonatal therapy and disease modeling. In this review we are aiming to summarize the knowledge on amniotic fluid stem cells and highlight the most promising results.
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Affiliation(s)
- Mara Cananzi
- Department of Paediatric Surgery, UCL Institute of Child Health & Great Ormond Street Hospital, London, UK
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