Isolation and Characterization of Novel Murine Epiphysis Derived Mesenchymal Stem Cells

Cheng, Chi‐Feng; Lian, Wei; Hsiao, Felix Shih-Hsiang; Liu, I-Hsuan; Lin, Shau-Ping; Lee, Yen-Hua; Chang, Chi Ching; Xiao, Guan-Yu; Huang, Hsin–Yi; Cheng, Ching‐Feng; Cheng, W. T. K.; Wu, Shinn-Chih

doi:10.1371/journal.pone.0036085

Cited by 33 publications

(36 citation statements)

References 75 publications

(81 reference statements)

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“…This loss of proliferation probably occurred because cellular senescence. Similarly, Cheng et al (2012) showed that MSC derived from mouse BM could be propagated until passage five, demonstrating a low proliferative potential of these cells. Although there is a decrease of cell proliferation rate at P3-4, these cells retained the differentiation potential.…”

Section: Discussionmentioning

confidence: 96%

Morphology and morphometry of feline bone marrow-derived mesenchymal stem cells in culture

et al. 2014

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RESUMO.-[Morfologia e morfometria das células-tronco mesenquimais isoladas de medula óssea de gato.] As células tronco mesenquimais são utilizadas na terapia de várias doenças na medicina humana e veterinária. As célu-las tronco foram isoladas da medula óssea de gato, entretanto, existem poucos dados referentes a morfologia e não existem informações sobre a morfometria das células tronco isoladas da medula óssea. Os objetivos do presente estudo foram o isolamento, avaliação do crescimento, potencial de diferenciação e caracterização morfológica e morfomé-trica das células mesenquimais de gato isoladas de medula óssea. A diferenciação in vitro foi realizada para confirmar a Mesenchymal stem cells (MSC) are increasingly being proposed as a therapeutic option for treatment of a variety of different diseases in human and veterinary medicine. Stem cells have been isolated from feline bone marrow, however, very few data exist about the morphology of these cells and no data were found about the morphometry of feline bone marrow-derived MSCs (BM-MSCs). The objectives of this study were the isolation, growth evaluation, differentiation potential and characterization of feline BM-MSCs by their morphological and morphometric characteristics. In vitro differentiation assays were conducted to confirm the multipotency of feline MSC, as assessed by their ability to differentiate into three cell lineages (osteoblasts, chondrocytes, and adipocytes). To evaluate morphological and morphometric characteristics the cells are maintained in culture. Cells were observed with light microscope, with association of dyes, and they were measured at 24, 48, 72 and 120h of culture (P1 and P3). The non-parametric ANOVA test for independent samples was performed and the means were compared by Tukey's test. On average, the number of mononuclear cells obtained was 12.29 (±6.05x10 6 ) cells/mL of bone marrow. Morphologically, BM-MSCs were long and fusiforms, and squamous with abundant cytoplasm. In the morphometric study of the cells, it was observed a significant increase in average length of cells during the first passage. The cell lengths were 106.97±38.16µm and 177.91±71.61µm, respectively, at first and third passages (24 h). The cell widths were 30.79±16.75 µm and 40.18±20.46µm, respectively, at first and third passages (24 h).The nucleus length of the feline BM-MSCs at P1 increased from 16.28µm (24h) to 21.29µm (120h). However, at P3, the nucleus length was 26.35µm (24h) and 25.22µm (120h). This information could be important for future application and use of feline BM-MSCs. ) células/mL de medula óssea. As células mesenquimais são longas e fusiformes, e escamosas com citoplasma abundante. No estudo morfométrico, observou-se aumento no comprimento médio das células durante a primeira passagem. As medidas de comprimento das células foram: 106,97±38,16µm e 177,91±71,61µm, respectivamente, na primeira e terceira passagens (24 horas). As medidas de largura das células foram: 30,79±16,75 µm e 40,18±20,46 µm, respectivamente, na primeira e terceir...

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Section: Discussionmentioning

confidence: 96%

Morphology and morphometry of feline bone marrow-derived mesenchymal stem cells in culture

et al. 2014

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“…Nevertheless, CD166 + has been described as an inconsistent marker, as this protein gradually decreases over many passages in murine epiphysis‐derived MSCs (Cheng et al. ). Whether this may be the case for rbMSCs remains to be elucidated, hence the loss of detection of this surface marker may not be enough to exclude the presence of rbMSCs.…”

Section: Discussionmentioning

confidence: 99%

Isolation, characterization and the multi‐lineage differentiation potential of rabbit bone marrow‐derived mesenchymal stem cells

et al. 2013

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Mesenchymal stem cells (MSCs) are recognized by their plastic adherent ability, fibroblastic-like appearance, expression of specific surface protein markers, and are defined by their ability to undergo multi-lineage differentiation. Although rabbit bone marrow-derived MSCs (rbMSCs) have been used extensively in previous studies especially in translational research, these cells have neither been defined morphologically and ultrastructurally, nor been compared with their counterparts in humans in their multi-lineage differentiation ability. A study was therefore conducted to define the morphology, surface marker proteins, ultrastructure and multi-lineage differentiation ability of rbMSCs. Herein, the primary rbMSC cultures of three adult New Zealand white rabbits (at least 4 months old) were used for three independent experiments. rbMSCs were isolated using the gradient-centrifugation method, an established technique for human MSCs (hMSCs) isolation. Cells were characterized by phase contrast microscopy observation, transmission electron microscopy analysis, reverse transcriptase-polymerase chain reaction (PCR) analysis, immunocytochemistry staining, flow cytometry, alamarBlue â assay, histological staining and quantitative (q)PCR analysis. The isolated plastic adherent cells were in fibroblastic spindle-shape and possessed eccentric, irregular-shaped nuclei as well as rich inner cytoplasmic zones similar to that of hMSCs. The rbMSCs expressed CD29, CD44, CD73, CD81, CD90 and CD166, but were negative (or dim positive) for CD34, CD45, CD117 and HLD-DR. Despite having similar morphology and phenotypic expression, rbMSCs possessed significantly larger cell size but had a lower proliferation rate as compared with hMSCs. Using established protocols to differentiate hMSCs, rbMSCs underwent osteogenic, adipogenic and chondrogenic differentiation. Interestingly, differentiated rbMSCs demonstrated higher levels of osteogenic (Runx2) and chondrogenic (Sox9) gene expressions than that of hMSCs (P < 0.05). There was, however, no difference in the adipogenic (Pparc) expressions between these cell types (P > 0.05). rbMSCs possess similar morphological characteristics to hMSCs, but have a higher potential for osteogenic and chondrogenic differentiation, despite having a lower cell proliferation rate than hMSCs. The characteristics reported here may be used as a comprehensive set of criteria to define or characterize rbMSCs.

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“…Regarding preparation of BM stromal responders, BM stromal cell (BMSC) cultures were set up from epiphyses from 5 mice per group, as described previously (46 Chimeric experiments. Mice were irradiated twice with an individual dose of 650 cGy with 4 hours recovery time between.…”

Section: V617fmentioning

confidence: 99%

IL-33 signaling contributes to the pathogenesis of myeloproliferative neoplasms

Mager¹,

Riether²,

Schürch³

et al. 2015

J. Clin. Invest.

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Isolation and Characterization of Novel Murine Epiphysis Derived Mesenchymal Stem Cells

Cited by 33 publications

References 75 publications

Morphology and morphometry of feline bone marrow-derived mesenchymal stem cells in culture

Morphology and morphometry of feline bone marrow-derived mesenchymal stem cells in culture

Isolation, characterization and the multi‐lineage differentiation potential of rabbit bone marrow‐derived mesenchymal stem cells

IL-33 signaling contributes to the pathogenesis of myeloproliferative neoplasms

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