Expression of OCT-4 and SOX-2 in Bone Marrow-Derived Human Mesenchymal Stem Cells during Osteogenic Differentiation

Matić, Igor; Antunović, Maja; Brkić, Šime; Josipović, Pavle; Mihalić, Katarina Caput; Karlak, Ivan; Ivković, Alan; Marijanović, Inga

doi:10.3889/oamjms.2016.008

Cited by 49 publications

(32 citation statements)

References 51 publications

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“…Further analysis of protein expression in subsequent passages could provide more information about the correlation between the gene and protein expression of the markers. Moreover, it is important to mention, that MSCs expanded in culture with bFGF were found to have downregulated SOX2 mRNA suggesting bFGF as a potential factor affecting the expression of transcriptional factors [116]. Therefore, defining a cell state and phenotype solely on transcript expression can be unreliable and protein analysis has to be included.…”

Section: Discussionmentioning

confidence: 99%

Differentiation of Human Mesenchymal Stem Cells from Wharton’s Jelly Towards Neural Stem Cells Using a Feasible and Repeatable Protocol

Kruminis-Kaszkiel

Osowski

Bejer-Oleńska

et al. 2020

Cells

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The transplantation of neural stem cells (NSCs) capable of regenerating to the cells of the central nervous system (CNS) is a promising strategy in the treatment of CNS diseases and injury. As previous studies have highlighted mesenchymal stem cells (MSCs) as a source of NSCs, this study aimed to develop a feasible, efficient, and reproducible method for the neural induction of MSCs isolated from Wharton's jelly (hWJ-MSCs). We induced neural differentiation in a monolayer culture using epidermal growth factor, basic fibroblast growth factor, N2, and B27 supplements. This resulted in a homogenous population of proliferating cells that expressed certain neural markers at both the protein and mRNA levels. Flow cytometry and immunocytochemistry confirmed the expression of neural markers: nestin, sex-determining region Y (SRY) box 1 and 2 (SOX1 and SOX2), microtubule-associated protein 2 (MAP2), and glial fibrillary acidic protein (GFAP). The qRT-PCR analysis revealed significantly enhanced expression of nestin and MAP2 in differentiated cells. This study confirms that it is possible to generate NSCs-like cells from hWJ-MSCs in a 2D culture using a practical method. However, the therapeutic effectiveness of such differentiated cells should be extended to confirm the terminal differentiation ability and electrophysiological properties of neurons derived from them.

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Section: Discussionmentioning

confidence: 99%

Differentiation of Human Mesenchymal Stem Cells from Wharton’s Jelly Towards Neural Stem Cells Using a Feasible and Repeatable Protocol

Kruminis-Kaszkiel

Osowski

Bejer-Oleńska

et al. 2020

Cells

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“…Ben-Shushan et al showed that the extinction of POU5F1 gene activity in stem cell-fibroblast hybrid cells was accompanied by fast methylation of regulatory sequences, such as proximal and distal enhancers in the POU5F1 promoter/enhancer region [45,46]. The expression of Oct4 and/or NANOG as markers of pluripotency was also demonstrated in BM-MSCs and in human multipotent adult SCs generated in vitro from adult human liver, heart, and BM [47,48] Previously, it was shown that DMSO can cause arrest of cells in the G1 phase, a process that is regulated by p21 and cyclin D1 cell cycle regulators [26][27][28]. In this study, DMSO and the process of freezing did not significantly change the expression of p21 and cyclin D1 genes in hAFSCs, indicating the applicability of this cryopreservation protocol.…”

Section: Discussionmentioning

confidence: 99%

Human amniotic fluid stem cells (hAFSCs) expressing p21 and cyclin D1 genes retain excellent viability after freezing with (dimethyl sulfoxide) DMSO

Aziz

Fardyazar

Pashaei-Asl

et al. 2019

Bosn J of Basic Med Sci

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Human amniotic fluid stem cells (hAFSCs) have features intermediate between embryonic and adult SCs, can differentiate into lineages of all three germ layers, and do not develop into tumors in vivo. Moreover, hAFSCs can be easily obtained in routine procedures and there is no ethical or legal limitations regarding their use for clinical and experimental applications. The aim of this study was to assess the effect of slow freezing/thawing and two different concentrations of DMSO (10% DMSO + 90% fetal bovine serum [FBS] and 5% DMSO + 95% FBS) on the survival of hAFSCs. hAFSCs were obtained from 5 pregnant women during amniocentesis at 16-22 weeks of gestation. The expression of pluripotency markers (Octamer-binding transcription factor 4 [Oct4] and NANOG) by reverse transcription polymerase chain reaction and cell surface markers (cluster of differentiation [CD31], CD44, CD45, and CD90) by flow cytometry was analyzed before and after the slow-freezing. Cell viability was assessed by trypan blue exclusion or MTT assay. Quantitative mRNA expression of Oct4, NANOG, cyclin D1 and p21 was determined by real-time PCR before and after the slow-freezing. Pluripotency of hAFSCs was confirmed by NANOG and POU5F1 (Oct4) gene expression before and after slow-freezing. All hAFSC cultures were positive for CD44 and CD90. A higher viability of hAFSCs was observed after freezing with 90% FBS + 10% DMSO. There was increased expression of NANOG and decreased expression of POU5F1 gene after freezing, compared to control cells (before freezing). DMSO and the process of freezing did not significantly change the expression of p21 and cyclin D1 genes in hAFSCs. Overall, our results indicate the applicability of slow-freezing and DMSO in cryopreservation of SCs.

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“…The presence of OCT-4 expression almost exclusively around the hemmule border assumes significance because the expression of OCT4 in bone marrow mesenchyme cells indicated osteogenic differentiation [39]. Similarly, the distribution of SOX2 scattered cells was predominantly found around the hemmule edges ( Figure S12).…”

Section: Individual Cells Stained By Stem Cell Antibodies In Hemmulementioning

confidence: 93%

Hemmule: A Novel Structure with the Properties of the Stem Cell Niche

Vodyanoy

Pustovyy

Globa

et al. 2020

IJMS

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Stem cells are nurtured and regulated by a specialized microenvironment known as stem cell niche. While the functions of the niches are well defined, their structure and location remain unclear. We have identified, in rat bone marrow, the seat of hematopoietic stem cells—extensively vascularized node-like compartments that fit the requirements for stem cell niche and that we called hemmules. Hemmules are round or oval structures of about one millimeter in diameter that are surrounded by a fine capsule, have afferent and efferent vessels, are filled with the extracellular matrix and mesenchymal, hematopoietic, endothelial stem cells, and contain cells of the megakaryocyte family, which are known for homeostatic quiescence and contribution to the bone marrow environment. We propose that hemmules are the long sought hematopoietic stem cell niches and that they are prototypical of stem cell niches in other organs.

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Expression of OCT-4 and SOX-2 in Bone Marrow-Derived Human Mesenchymal Stem Cells during Osteogenic Differentiation

Cited by 49 publications

References 51 publications

Differentiation of Human Mesenchymal Stem Cells from Wharton’s Jelly Towards Neural Stem Cells Using a Feasible and Repeatable Protocol

Differentiation of Human Mesenchymal Stem Cells from Wharton’s Jelly Towards Neural Stem Cells Using a Feasible and Repeatable Protocol

Human amniotic fluid stem cells (hAFSCs) expressing p21 and cyclin D1 genes retain excellent viability after freezing with (dimethyl sulfoxide) DMSO

Hemmule: A Novel Structure with the Properties of the Stem Cell Niche

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