Characterization of Equine Adipose Tissue‐Derived Stromal Cells: Adipogenic and Osteogenic Capacity and Comparison with Bone Marrow‐Derived Mesenchymal Stromal Cells

Vidal, Martin A.; Kilroy, Gail; Lopez, Mandi J.; Johnson, Jill; Moore, Rustin M.; Gimble, Jeffrey M.

doi:10.1111/j.1532-950x.2007.00313.x

Cited by 203 publications

(239 citation statements)

References 36 publications

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“…Vidal et al (2007) also observed that cell populations obtained from adipose tissue did not form as many colonies as cells from bone marrow, and that they were distributed more uniformly and adhered more firmly to the culture plate, as observed in this study.…”

Section: Discussionsupporting

confidence: 65%

“…The cell population with the fusiform appearance described here in samples from both adipose tissue and bone marrow were morphologically similar to mesenchymal stem cells from human (Zuk et al 2001, You et al 2009), sheep (Fadel et al 2011), horse (Fortier et al 1998, Smith et al 2003, Hegewald et al 2004, Vidal et al 2006, 2007, Maia et. al 2012, De Vita et al 2013, and other animals reported in many studies, in which these cells were described as star-shaped or fibroblast-like.…”

Section: Discussionmentioning

confidence: 76%

“…In veterinary medicine, knowledge of these cells is still very limited, and they are utilized based on the hypothesis that their essential characteristics, such as time of multiplication and potential for differentiation, are the same for all species (Vidal et al 2007). However, according to Fraser et al (2008), stem cells from adipose tissue and from bone marrow of the same individual are similar but not identical.…”

Section: Introductionmentioning

confidence: 99%

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Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

2013

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Pesq. Vet. Bras. 33(Supl.1):20-24, dezembro 2013 20 RESUMO.-[Cultivo de células da fração mononuclear da medulla óssea e da fração vascular stromal do tecido adipose de equinos em diferentes meios.] O objetivo deste estudo foi avaliar o cultivo de células da fração mononuclear da medula óssea e da fração vascular estromal do tecido adiposo de equinos em dois diferentes meios. Cinco cavalos foram submetidos à aspiração da medula óssea do esterno e à coleta de tecido adiposo da região glútea, pró-xima à inserção da cauda. A fração mononuclear e a fração vascular estromal foram obtidas das amostras e cultivadas em meio DMEM suplementado com soro fetal bovino a 10% ou em meio AIM-V. As culturas foram observadas uma vez por semana com um microscópio de luz invertida, com o intuito de se realizar uma análise qualitativa das características morfológicas das células, bem como do aspecto geral do cultivo celular. As unidades formadoras de colônia (CFU) foram contadas nos dias 5, 15 e 25 do cultivo celular. Durante a primeira semana, foram observadas diferenças entre amostras obtidas de mesma origem mantidas em diferentes meios. O número de colônias foi significativamente maior nas amostras de medula óssea em relação às amostras de tecido adiposo.TERMOS DE INDEXAÇÃO: Cavalos, medula óssea, tecido adiposo, meios de cultura. INTRODUCTIONThe bone marrow aspirate concentrate (BMAC) has been used to induce bone formation in humans with osseous defects, nonunion, or osteonecrosis. In horses BMAC has been used to improve cartilage, bone, and tendon repair, The objective of this study was to evaluate the culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in two different cell culture media. Five adult horses were submitted to bone marrow aspiration from the sternum, and then from the adipose tissue of the gluteal region near the base of the tail. Mononuclear fraction and stromal vascular fraction were isolated from the samples and cultivated in DMEM medium supplemented with 10% fetal bovine serum or in AIM-V medium. The cultures were observed once a week with an inverted microscope, to perform a qualitative analysis of the morphology of the cells as well as the general appearance of the cell culture. Colony-forming units (CFU) were counted on days 5, 15 and 25 of cell culture. During the first week of culture, differences were observed between the samples from the same source maintained in different culture media. The number of colonies was significantly higher in samples of bone marrow in relation to samples of adipose tissue.

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Section: Discussionsupporting

confidence: 65%

Section: Discussionmentioning

confidence: 76%

Section: Introductionmentioning

confidence: 99%

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Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

2013

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“…For the subsequent passages, cells were inoculated in 5 cm 2 flasks at 25!10 3 cells/cm 2 and allowed to multiply for 6-7 days to 90% confluence before trypsinization and successive passage. DT and CD were calculated from haemocytometer counts and cell culture time (CT) for each passage according to the following two formulae (Vidal et al 2007):…”

Section: Cell Doubling Methodsmentioning

confidence: 99%

Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

Iacono

Brunori²,

Pirrone³

et al. 2012

Reproduction

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Mesenchymal stem cells (MSCs) have been derived from multiple sources of the horse including umbilical cord blood (UCB) and amnion. This work aimed to identify and characterize stem cells from equine amniotic fluid (AF), CB and Wharton's Jelly (WJ). Samples were obtained from 13 mares at labour. AF and CB cells were isolated by centrifugation, while WJ was prepared by incubating with an enzymatic solution for 2 h. All cell lines were cultured in DMEM/TCM199 plus fetal bovine serum. Fibroblast-like cells were observed in 7/10 (70%) AF, 6/8 (75%) CB and 8/12 (66.7%) WJ samples. Statistically significant differences were found between cell-doubling times (DTs): cells isolated from WJ expanded more rapidly (2.0G0.6 days) than those isolated from CB (2.6G1.3 days) and AF (2.3G1.0 days) (P!0.05). Positive von Kossa and Alizarin Red S staining confirmed osteogenesis. Alcian Blue staining of matrix glycosaminoglycans illustrated chondrogenesis and positive Oil Red O lipid droplets staining suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105; and negative for CD34, CD14 and CD45. These findings suggest that equine MSCs from AF, UCB and WJ appeared to be a readily obtainable and highly proliferative cell lines from a uninvasive source that may represent a good model system for stem cell biology and cellular therapy applications in horses. However, to assess their use as an allogenic cell source, further studies are needed for evaluating the expression of markers related to cell immunogenicity.

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“…Stem cells are an interesting cellular model to study differentiation mechanisms because of the relative ease of establishing in vitro cultures and their good proliferation (Bianco et al 2001). Equine mesenchymal stem cells have been established in vitro from different tissues such as bone marrow (Fortier et al 1998), umbilical cord blood (Reed and Johnson 2008) and adipose tissue (Vidal et al 2007) and show differentiation capabilities into mesodermal lineages. Nevertheless, alternative cell sources to be used in veterinary clinical applications such as tissue engineering and repair are still necessary.…”

Section: Introductionmentioning

confidence: 99%

Isolation and differentiation potential of an equine amnion-derived stromal cell line

et al. 2011

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Stem cells represent an important tool in veterinary therapeutic field such as tissue engineering. In the present study, equine amnion-derived mesenchymal stromal cells were investigated for applications in veterinary science as an alternative source to bone marrow mesenchymal stem cells and adipose stem cells. Amnion stromal cells isolation and characterization protocol is described; the in vitro cell growth rate was calculated by measuring viable cell number over 20 days. The expression of stem cell markers such as Oct-4, Nanog, Sox-2 and CD105 was assessed by retrotranscription quantitative PCR (RT-qPCR) and differentiation into adipocytes, osteocytes and chondrocytes precursors was analyzed by cytochemical staining. This study showed that amnion stromal cells expressing stem cell markers can differentiate into mesoderm lineage and may be an alternative source to mesenchymal stem cells derived from adipose tissue and bone marrow for the use in tissue repair.

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Characterization of Equine Adipose Tissue‐Derived Stromal Cells: Adipogenic and Osteogenic Capacity and Comparison with Bone Marrow‐Derived Mesenchymal Stromal Cells

Abstract: Adipose tissue is a potential source of adult stem cells for tissue engineering applications in equine veterinary medicine.

Cited by 203 publications

References 36 publications

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

Isolation and differentiation potential of an equine amnion-derived stromal cell line

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