Protocols for obtainment and isolation of two mesenchymal stem cell sources in sheep

Fadel, Leandro; Viana, Brunno Rosa; Feitosa, Matheus Levi Tajra; Ercolin, Anna Carolina Mazeto; Roballo, Kelly Cristine Santos; Casals, Juliana Barbosa; Pieri, Naira Caroline Godoy; Meirelles, Flávio Vieira; Martins, Daniele dos Santos; Miglino, María Angélica; Ambrósio, Carlos Eduardo

doi:10.1590/s0102-86502011000400004

Cited by 45 publications

(32 citation statements)

References 26 publications

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“…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: Discussionsupporting

confidence: 63%

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: 63%

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

2013

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“…However, harvesting MSCs from heterogenous cell population still remains one of the challenging tasks that are worth considering (Fekete et al, 2012;Harichandan and Buhring, 2011;Hsu et al, 2012). Despite of several advancement and sophistication for MSC isolation was made in case of human and small laboratory research animal, a standard and convenient method of isolating MSCs from large domestic animal (Fadel et al, 2011;Stewart et al, 2007) still need to be optimized. De Schauwer et al (2014) reported the non-invasive sources of MSCs such as umbilical cord blood, umbilical cord matrix and peripheral blood.…”

Section: Introductionmentioning

confidence: 99%

A Comparative Analysis of Invasive and Non-Invasive Method of Bone Marrow Stromal Cell Isolation

Santra¹,

Gupta²,

Singh³

et al. 2015

Asian J. of Animal and Veterinary Advances

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The conventional method of Bone Marrow Stromal Cell (BMSC) isolation from live subjects are complex due to involvement of lot of expert personnel and pre and post operational medication and cares. Moreover, the concerning ethical issues also pose lots of restrictions for isolation of BMSC's making stem cells research restricted to certain elite laboratories only. This study aims to compare between the regular aspiration (invasive) method and an alternative, straight forward and non-invasive method of BMSC harvest. The BMSCs were harvested by both invasive and non-invasive methods and cultured in MSC (Mesenchymal Stem Cell) medium. The cells were undergone visual assessment of growth dynamics as well as identification and characterization of MSC cells, based on microscopic examination. Both of these tested methods successfully yielded significant amount of BMSC that were found to be identical in morphology, growth dynamics and in vitro cultural properties. Unlike the invasive method that requires a live animal, the non-invasive method relies on post-slaughtered bone and therefore obviates the requirement of skill personnel and setup. Eventually, the used bone from already dead animal no longer becomes the issue of conflict with animal ethics and welfare. The ease in cell harvest and lack of ethical barrier would definitely make BMSC harvest convenient and therefore, anticipated to be well accepted in resource poor laboratories around the globe.

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“…Regarding such animals, outcomes were found in studies about neurodegenerative injuries 12,13 , as well as in attempts to use cell therapy as a way to treat cardiovascular diseases 20 . Experiments with sheep were described for obtaining the main uses of stem cells/progenitor cells 21,22 . However, such potentialities have been little studied in the goat species.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

Filho¹,

Neto²,

Carvalho³

et al. 2014

Acta Cir. Bras.

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PURPOSE:To characterize bone marrow progenitors cells grown in vitro, using native goats from northeastern Brazil as animal model. METHODS:Ten northeastern Brazil native goats of both genders were used from the Piauí Federal University Agricultural Science Center's (UFPI) -Goat Farming Sector. Bone marrow aspirates where taken from the tibial ridge and seeded on culture plates for isolation, expansion and Flow Cytometry (expression markers -Oct-3/4, PCNA, Ck-Pan, Vimentina, Nanog). RESULTS:Progenitor cells showed colonies characterized by the presence of cell pellets with fibroblastoid morphology. Cell confluence was taken after 14 days culture and the non-adherent mononuclear cell progressive reduction. After the first passage, 94.36% cell viability was observed, starting from 4.6 x 10 6 cell/mL initially seeded. Cells that went through flow cytometry showed positive expression for Oct-3/4, PCNA, Ck-Pan, Vimentina, and Nanog. CONCLUSIONS:Bone marrow progenitor isolated of native goats from northeastern Brazil showed expression markers also seen in embryonic stem cells (Oct-3/4, Nanog), markers of cell proliferation (PCNA) and markers for mesenchymal cells (Vimentina and Ck-pan), which associated to morphological and culture growth features, suggest the existence of a mesenchymal stem cell (MSC) population in the goat bone marrow stromal cells studied.Key words: Cell Culture Techniques. Cell Differentiation. Stem Cells. Flow Cytometry. Goats. Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

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Protocols for obtainment and isolation of two mesenchymal stem cell sources in sheep

Cited by 45 publications

References 26 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

A Comparative Analysis of Invasive and Non-Invasive Method of Bone Marrow Stromal Cell Isolation

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

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