Osteogenic proliferation and differentiation of canine bone marrow and adipose tissue derived mesenchymal stromal cells and the influence of hypoxia

Chung, Dai-Jung; Hayashi, Kei; Toupadakis, Chrisoula A.; Wong, Alice; Yellowley, Clare E.

doi:10.1016/j.rvsc.2010.10.012

Cited by 58 publications

(63 citation statements)

References 54 publications

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“…Osteogenesis is tightly coupled with angiogenesis during bone development and regeneration. It has been reported that over-expression of Cbfα1 in fibroblasts induces VEGF production by stimulating VEGF transcription (Zelzer et al 2001;Chung et al 2010). In the present study, we also showed that at the later stage of hypoxia exposure, Cbfα1 expression was increased, but whether VEGF is involved in the differentiation of MSCs into osteogenic lineages remains to be investigated.…”

Section: Discussionsupporting

confidence: 67%

Hypoxia Induces Osteogenesis-Related Activities and Expression of Core Binding Factor .ALPHA.1 in Mesenchymal Stem Cells

Huang

Deng

Wang

et al. 2011

Tohoku J. Exp. Med.

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Mesenchymal stem sells (MSCs) have received much attention in the field of bone tissue engineering due to their biological capability to differentiate into osteogenic lineage cells. Hypoxia-inducible factor 1alpha (HIF-1α) plays an important role in the MSC-related bone regeneration during hypoxia, while core binding factor alpha 1 (Cbfα1) is a transcription regulator that is involved in the chondrocyte differentiation and ossification. In the present study, we investigated the effects of hypoxia on biological capability of MSCs. MSCs were isolated from adult rabbit bone marrow, and were cultured in vitro under normoxia (air with 5% CO 2 ) or hypoxia (5% CO 2 and 95% N 2 ). The proliferation of MSCs, alkaline phosphatase (ALP) activity, and production of collagens type I and type III (Col I/III) were examined. The expression levels of HIF-1α and Cbfα1 were measured by real-time PCR and western blot analyses. We found that hypoxia significantly induced the proliferation of MSCs and increased ALP activity and the production of Col I/III. Moreover, hypoxia increased the expression of Cbfα1 mRNA after 12 h, whereas the expression of HIF-1α mRNA was increased after 1 h of hypoxia. Knockdown of HIF-1α expression with a small interfering RNA significantly increased the expression levels of Cbfα1 protein either under the normoxia or hypoxia condition. Our results indicate that hypoxia enhances MSCs to differentiate into osteogenic lineage cells and suggest that Cbfα1 may be negatively regulated by HIF-1α.

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Section: Discussionsupporting

confidence: 67%

Hypoxia Induces Osteogenesis-Related Activities and Expression of Core Binding Factor .ALPHA.1 in Mesenchymal Stem Cells

Huang

Deng

Wang

et al. 2011

Tohoku J. Exp. Med.

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“…MSCs were isolated from fat and cultured exactly as previously described for canine MSCs [21][22][23]. MSCs were cultured in low-glucose Dulbecco's modified Eagle's medium (Mediatech), 10% fetal bovine serum (HyClone, Inc.), and 1% penicillin/streptomycin (Thermo Fisher Scientific) in tissue culture flasks (Nunc) and incubated at 37°C, 5% CO 2 .…”

Section: Fmsc Isolation and Expansionmentioning

confidence: 99%

Feline Foamy Virus Adversely Affects Feline Mesenchymal Stem Cell Culture and Expansion: Implications for Animal Model Development

Arzi

Kol

Murphy

et al. 2015

Stem Cells and Development

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Mesenchymal stem cells (MSCs) are a promising therapeutic option for various immune-mediated and inflammatory disorders due to their potent immunomodulatory and trophic properties. Naturally occurring diseases in large animal species may serve as surrogate animal models of human disease, as they may better reflect the complex genetic, environmental, and physiologic variation present in outbred populations. We work with naturally occurring diseases in large animal species to better understand how MSCs work and to facilitate optimal translation of MSC-based therapies. We are investigating the use of MSC therapy for a chronic oral inflammatory disease in cats. During our efforts to expand fat-derived feline MSCs (fMSCs), we observed that*50% of the cell lines developed giant foamy multinucleated cells in later passages. These morphologic alterations were associated with proliferation arrest. We hypothesized that the cytopathic effects were caused by infection with a retrovirus, feline foamy virus (FFV). Using transmission electron microscopy, polymerase chain reaction, and in vitro assays, we determined that syncytial cell formation and proliferation arrest in fMSCs were caused by FFV strains that were highly homologous to previously reported FFV strains. We determined that the antiretroviral drug, tenofovir, may be used to support ex vivo expansion and salvage of FFV-infected fMSC lines. MSC lines derived from specific pathogen-free cats do not appear to be infected with FFV and may be a source of allogeneic fMSCs for clinical application. FFV infection of fMSC lines may hinder large-scale expansion of autologous MSC for therapeutic use in feline patients.

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“…Após a diferenciação osteogênica as CTM adquirem forma cuboidal ou poligonal semelhante aos osteoblastos e se agrupam concentricamente formando nódulos mineralizados (Eslaminejad & Taghiyar 2010). Esses nódulos são considerados indicadores de diferenciação osteogênica "in vitro" (Chung et al 2012, Spencer et al 2012) e sua quantificação é a forma mais fidedigna de avaliar a osteogênese "in vitro", visto que a produção de matriz mineralizada, não indica apenas a fase final da diferenciação osteogênica, mas também a atividade metabólica das células diferenciadas (Bruedigam et al 2011).…”

Section: Discussionunclassified

“…Ela é expressa por osteoblastos e reflete também o final da diferenciação e da maturação da matriz óssea e é considerada o marcador tardio mais específico da diferenciação osteogênica (Nakashima et al 2002). O aumento crescente de sua expressão é caracterís-tico do processo de diferenciação osteogênica de CTM-MO e CTM-AD (Chung et al 2012). A osteonectina também está relacionada com a mineralização da matriz no estágio final de diferenciação (Nakashima et al 2002).…”

Section: Discussionunclassified

“…Muitos estudos já foram realizados em humanos e ratos mas, pouco se sabe sobre o potencial osteogênico dessas células em outras espé-cies como o cão. Alguns pesquisadores demonstram que as CTM oriundas da medula óssea (CTM-MO) apresentam maior potencial osteogênico que as CTM do tecido adiposo (CTM-AD) , Monaco et al 2012, outros indicam maior potencial osteogênico nas CTM-AD (Lin et al 2009, Chung et al 2012) e existem aqueles que mostram potencial osteogênico semelhante entre os dois tipos celulares (Spencer et al 2012, Ranera et al 2012. Em cães, os estudos comparativos entre CTM-MO e CTM-AD são escassos.…”

Section: Introductionunclassified

See 1 more Smart Citation

Estudo comparativo da diferenciação osteogênica das células tronco mesenquimais da medula óssea e do tecido adiposo de cães adultos

et al. 2016

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Resumo: O objetivo deste estudo foi comparar o potencial osteogênico das células tronco mesenquimais extraídas da medula óssea (CTM-MO) com as do tecido adiposo (CTM-AD) de cães adultos. As células foram caracterizadas fenotipicamente quanto à expressão de CD29, CD90, CD34 e CD45 e submetidas à diferenciação adipogênica e condrogênica por 21 dias e osteogênica por 7, 14 e 21 dias. Foram constituídos quatro grupos: 1) CTM-MO em meio osteogênico, 2) CTM-MO em meio basal, 3) CTM-AD em meio osteogênico e 4) CTM-AD em meio basal. Aos 7, 14 e 21 dias de diferenciação osteogênica as culturas foram submetidas às avaliações da conversão de MTT em formazan, da atividade da fosfatase alcalina (FA), da síntese de colágeno e de matriz mineralizada, avaliação do número de células por campo e foram quantificados os transcritos gênicos para osterix, sialoproteina óssea (BSP), osteonectina (ON) e osteocalcina (OC). Tanto as células extraídas da medula óssea quanto do tecido adiposo mostraram elevada expressão de marcadores para células tronco e baixa expressão de marcadores de células hematopoiéticas (menor que 2%). Além disso, foram capazes de se diferenciar em osteoblastos, condrócitos e adipócitos. As CTM-AD submetidas à diferenciação osteogênica mostraram maior conversão do MTT em formazan que as CTM-MO, sob mesmas condições aos 7 e 21 dias. O número de células por campo, a atividade da FA, a síntese de colágeno e de matriz mineralizada foram superior nas CTM-AD em diferenciação, em relação às CTM-MO sob as mesmas condições, em todos os tempos estudados. As expressões de osterix, BSP e OC foram predominantemente superiores nas CTM-MO diferenciadas, mas a expressão de ON foi superior nas CTM-AD diferenciadas aos 7, 14 e 21 dias. Conclui-se que as CTM-AD apresentam maior potencial osteogênico que as CTM-MO quando extraídas de cães adultos.

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Osteogenic proliferation and differentiation of canine bone marrow and adipose tissue derived mesenchymal stromal cells and the influence of hypoxia

Cited by 58 publications

References 54 publications

Hypoxia Induces Osteogenesis-Related Activities and Expression of Core Binding Factor .ALPHA.1 in Mesenchymal Stem Cells

Hypoxia Induces Osteogenesis-Related Activities and Expression of Core Binding Factor .ALPHA.1 in Mesenchymal Stem Cells

Feline Foamy Virus Adversely Affects Feline Mesenchymal Stem Cell Culture and Expansion: Implications for Animal Model Development

Estudo comparativo da diferenciação osteogênica das células tronco mesenquimais da medula óssea e do tecido adiposo de cães adultos

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