Fetal mesenchymal stem cells derived from human umbilical cord sustain primitive characteristics during extensive expansion

Jo, Chris Hyunchul; Kim, Ok Su; Park, Eun Young; Kim, Byoung Jae; Lee, Ji Ho; Kang, Sungchul; Lee, Jae Hyup; Han, Hyuck; Rhee, Seung Hwan; Yoon, Kang Sup

doi:10.1007/s00441-008-0696-3

Cited by 97 publications

(83 citation statements)

References 39 publications

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“…Go et al (18) reported that MSCs lost their stem cell characteristics after several generations but would maintain the potential for the proliferation and differentiation through transduction Sox2 or Nanog gene into the MSCs. And the levels of OCT4, Nanog, Sox2, and TERT mRNA expression correlated positively with the undifferentiated state, suggesting that OCT4, Nanog, Sox2, and TERT mRNA could be used as a sign of undifferentiated MSCs (18,19). Greco et al (12) found that these embryonic transcription factors could also be indicators of MSCs differentiation potential as an experimental tool and also in clinical diagnostics.…”

Section: Discussionmentioning

confidence: 99%

Effect of Recombinant Human Erythropoietin On the Stemness of Bone Marrow-derived Mesenchymal Stem Cells in vitro

Chen

et al. 2010

Int J Stem Cells

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The purpose of this study was to investigate the effects of the recombinant human erythropoietin (rhEPO) on proliferative and multi-differentiation potential of the bone marrow-derived mesenchymal stem cells (MSCs). The MSCs were isolated primarily from bone marrow of adult rat and purified at increasing passage. A purified population of MSCs can be obtained about 2 weeks after the initiation of culture. After three passages (P3-MSCs), bone marrow-derived adherent cells were identified, then different concentrations of rhEPO (0.1, 1, 5, 10, 100 U/ml) was added into the Passage-3 cells which had been identified. The expression of the surface markers in adherent cells was detected by the flow cytometry. The mRNA levels of transcription factors OCT4, SOX2, Nanog and TERT were measured by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that CD29 and CD90 were positive in MSCs, but not CD33, CD44 and CD45, and the cells could differentiate into multiple lineages such as osteocytes and adipocytes. The expression of OCT4, SOX2, TERT, Nanog mRNA were up-regulated by the treatment of EPO. The effect of EPO was the most obvious when its concentration was 5U/mL after 12h. we conclude that MSCs can not only perserve characteristics of stem cells but also maintain its multi-lineage differentiation potential after appropriate treatment of EPO.

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Section: Discussionmentioning

confidence: 99%

Effect of Recombinant Human Erythropoietin On the Stemness of Bone Marrow-derived Mesenchymal Stem Cells in vitro

Chen

et al. 2010

Int J Stem Cells

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“…However, in certain degenerative diseases, such as osteoarthritis, MSCs are depleted and have reduced ability to differentiate and proliferate [6] . As an alternative source of MSCs, young cells such as fetal (f) MSCs could be harvested from the umbilical cord, placental stroma, and the amnion [7][8][9] . Ethical concerns regarding the isolation of MSCs from fetal sources are minimized, as these tissues are normally discarded after birth [1] .…”

Section: Introductionmentioning

confidence: 99%

Fetal vs adult mesenchymal stem cells achieve greater gene expression, but less osteoinduction

Santiago-Torres

Lovasz

Bertone

2015

WJSC

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AIM:To investigate adenoviral transduction in mesenchymal stem cells (MSCs) and effects on stemness in vitro and function as a cell therapy in vivo . METHODS:Bone marrow-derived adult and fetal MSC were isolated from an equine source and expanded in monolayer tissue culture. Polyethylenimine (PEI)-mediated transfection of pcDNA3-eGFP or adenoviral transduction of green fluorescent protein (GFP) was evaluated in fetal MSCs. Adenoviral-mediated transduction was chosen for subsequent experiments. All experiments were carried out at least in triplicate unless otherwise noted. Outcome assessment was obtained by flow cytometry or immunohystochemistry and included transduction efficiency, cell viability, stemness (i.e. , cell proliferation, osteogenic and chondrogenic cell differentiation), and quantification of GFP expression. Fetal and adult MSCs were then transduced with an adenoviral vector containing the gene for the bone morphogenic protein 2 (BMP2). In vitro BMP2 expression was assessed by enzyme linked immunosorbent assay. In addition, MSC-mediated gene delivery of BMP2 was evaluated in vivo in an osteoinduction nude mouse quadriceps model. New bone formation was evaluated by microradiography and histology.RESULTS: PEI provided greater transfection and viability in fetal MSCs than other commercial chemical reagents. Adenoviral transduction efficiency was superior to PEI-mediated transfection of GFP in fetal MSCs (81.3% ± 1.3% vs 35.0% ± 1.6%, P < 0.05) and was similar in adult MSCs (78.1% ± 1.9%). Adenoviral transduction provided significantly greater expression of GFP in fetal than adult MSCs (7.4 ± 0.1 vs 4.4 ± 0.3 millions of mean fluorescence intensity units, P < 0.01) as well as significantly greater in vitro BMP2 expression (0.16 pg/cell-day vs 0.10 pg/cell-day, P < 0.01). Fraction of fetal MSC GFP positive cells decreased significantly faster than adult MSCs (1.15% ± 0.05% vs 11.4% ± 2.1% GFP positive at 2 wk post-transduction, P < 0.05). Cell proliferation and osteogenic differentiation in vitro Clinical Trials Studywere not affected by Ad transduction in both fetal and adult MSCs, but fetal MSCs had reduced chondrogenic differentiation in vitro when compared to adult (P < 0.01). Chondrogenic differentiation was also significantly reduced in Ad-GFP transduced cells (P < 0.05). Ad-BMP2 transduced adult MSCs induced new bone formation in more thighs than Ad-BMP2 transduced fetal MSCs (83% vs 17% of the six treated thighs per group, P < 0.05) and resulted in increased femur midshaft diameter due to greater extent of periosteal new bone (1.57 ± 0.35 mm vs 1.27 ± 0.08 mm, P < 0.05). CONCLUSION:Fetal MSCs may be genetically manipulated ex vivo with adenoviral vectors. Nonetheless, the abbreviated expression of the exogenous gene may limit their applications in vivo .

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“…Jo et al, [76] have shown that pluripotent stem cell markers, such as Oct-4, SSEA-3, SSEA-4, Tra-1-60, and Tra-1-81, are expressed, as both mRNA and protein, by MSCs from the whole cord and their expression levels are maintained up to the 9 th passage. Furthermore, Qiao et al, [75] have detected nucleostemin, a p52 binding protein localized in nucleoli of stem cells [82], and BMI-1 involved in self-renewal [83], whereas La Rocca et al, [77] have demonstrated, for the first time, the presence of both isoforms of Oct-4 (A and B).…”

Section: Mscs From Whole Umbilical Cordmentioning

confidence: 99%

“…Also in this case, the incubation times greatly differ from one laboratory to another, ranging from 10 min to 3 h. The most frequently used culture medium is DMEM-LG supplemented with 10% FBS or FCS. However, growth factors, such as vascular endothelial growth factor (VEGF), EGF, and bFGF, have been also added [72,76,79]. A comparison between the explant method and enzymatic digestion was carried out by Schugar et al, [81], which demonstrated that the isolation method affects population phenotype.…”

Section: Mscs From Whole Umbilical Cordmentioning

confidence: 99%

“…After mechanical chopping, explant culture of the cord is carried out or UC fragments are incubated at 37°C with various types of collagenase (0.1% or 0.075%, w/v) and trypsin (0.25% or 0.125%, w/v) [2,4,[72][73][74][75][76][77][78][79][80]. Also in this case, the incubation times greatly differ from one laboratory to another, ranging from 10 min to 3 h. The most frequently used culture medium is DMEM-LG supplemented with 10% FBS or FCS.…”

Section: Mscs From Whole Umbilical Cordmentioning

confidence: 99%

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Phenotype and Differentiation Potential of Stromal Populations Obtained from Various Zones of Human Umbilical Cord: An Overview

Conconi

Liddo²,

Tommasini³

et al. 2011

TOTERMJ

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Fibroblast-like cells with properties similar to mesenchymal stromal cells (MSCs) are present in human umbilical cord (hUC). In accordance with the international minimal criteria for defining multipotent mesenchymal stromal cells, hUC cells are designed MSCs being plastic adherent, positive to specific non hematopoietic lineage biomarkers, able to be both in vitro long term cultured and differentiated into osteoblasts, chondroblasts and adipocytes. In this review, a panoramic view of phenotypic characteristics of hUC cells derived from various UC parts are described. The high heterogeneity of extraction, culture and analysis procedures hinder the ability to precisely identify UC stromal cells. As a result, different phenotypic profiles are detectable not only among the cells obtained from the various parts of cord, but also inside the same UC regions, suggesting that UC-MSCs may represent an unique cell family whose components present various degree of stemness. However, in vitro and in vivo evidence indicates Wharton's jelly as the best source of MSCs, because its cells present a wide range of potential therapeutic applications.

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Fetal mesenchymal stem cells derived from human umbilical cord sustain primitive characteristics during extensive expansion

Cited by 97 publications

References 39 publications

Effect of Recombinant Human Erythropoietin On the Stemness of Bone Marrow-derived Mesenchymal Stem Cells in vitro

Effect of Recombinant Human Erythropoietin On the Stemness of Bone Marrow-derived Mesenchymal Stem Cells in vitro

Fetal vs adult mesenchymal stem cells achieve greater gene expression, but less osteoinduction

Phenotype and Differentiation Potential of Stromal Populations Obtained from Various Zones of Human Umbilical Cord: An Overview

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