Canine adipose-derived-mesenchymal stem cells do not lose stem features after a long-term cryopreservation

Martinello, Tiziana; Bronzini, Ilaria; Maccatrozzo, Lisa; Mollo, Antonio; Sampaolesi, Maurilio; Mascarello, Francesco; Decaminada, M; Patruno, Marco

doi:10.1016/j.rvsc.2010.07.024

Cited by 122 publications

(114 citation statements)

References 23 publications

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“…The cells that generated these clusters possibly were committed mesenchymal progenitors. Generally, in our culture system adipogenesis was weakly induced, but this was also previously observed during induction of adipogenesis in canine adipose tissue MSCs, possibly due to the type of medium used [24]. On the other hand, according to hierarchical model of in vivo and in vitro bone marrow mesenchymal progenitor differentiation, adipogenic potential is the fi rst to be lost and osteogenic is the last [25].…”

Section: Discussionmentioning

confidence: 58%

Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

et al. 2015

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The aim of this study was to determine whether synovial fl uid (SF) of dogs contains cells that have characteristics of MSCs and to describe their differentiation potential. SF adherent cells from 5 young German shepherd dogs (average 3.8 ± 0.9 years) were expanded (37ºC, 5% CO 2 , humidifi ed atmosphere) three weeks before their phenotype was characterized by fl ow-cytometry for the presence of CD90 and CD34. Population doubling time (PDT), number of CFU-F and adipogenic, osteogenic and chondrogenic potentials have been determined in vitro. In early passages PTD was 31 ± 10 hours and expansion fold after 3 sub cultivations (9 days) theoretically could be 372 ± 134. At P1, 0.55 ± 0.05% of SF cells had the ability to form CFU-F. Sixty-six percent of cells expressed CD90 and none of the cells expressed markers of hematopoietic cells. Oil Red O staining has shown accumulation of fat droplets in cells grown in adipogenic medium, while deposits of calcium in the osteogenic medium were evidenced with Alizarin red staining. SF cultured in hondrogenic and control medium in three-dimensional conditions formed a cartilage-like tissue. Alcian blue staining of pellets' slides have shown a signifi cant amount of glycosaminoglycans (GAGs) and immunohistochemistry analysis documented collagen type II expression. The amount of GAGs in pellets grown in both conditions showed no difference. SF cells in vitro exhibited osteogenic, adipogenic and chondrogenic differentiation potentials, suggesting the presence of different mesenchymal progenitors. These results also demonstrated that SF cells have a spontaneous chondrogenic potential that should be further explored for possible tissue engineering protocols.

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Section: Discussionmentioning

confidence: 58%

Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

et al. 2015

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“…A alta expressão de marcadores de células tronco e a baixa expressão de marcadores de células hematopoiéticas observadas no presente estudo, confirmam o sucesso no isolamento das CTM a partir de amostras de medula óssea e de tecido adiposo. Esse padrão de expressão é mundialmente aceito como característico das células tronco mesenquimais (Dominici et al 2006, Aksu et al 2008, Levi et al 2011, Martinello et al 2011. A International Society for Cellular Therapy definiu que as CTM devem expressar marcadores como CD105, CD73 e CD90 e não expressar os marcadores de células hematopoiéticas (Dominici et al 2006).…”

Section: Discussionunclassified

“…De forma semelhante a elevada expressão de CD29, marcador de CTM não é expresso por fibroblastos caninos , corrobora com o isolamento de CTM e exclui a possibilidade do isolamento de fibroblastos, principalmente das culturas extraídas do tecido adiposo. Outros marcadores de CTM podem também ser utilizados em cães como CD44 (Vieira et al 2010), CD33, CD105, CD184, OCT4 (Seo et al 2009), CD140a, CD117 (Martinello et al 2011), CD56, CD146, CD271 (Rozemuller et al 2010), mas não haviam, na época do experimento, anticorpos específicos para cão disponíveis no mercado. A menor conversão de MTT em cristais de formazan observada nas CTM submetidas à diferenciação osteogênica no presente estudo quando comparadas às CTM cultivadas em meio basal, sugerem menor atividade mitocondrial nos grupos submetidos à diferenciação, o que também foi observado em CTM de ratos (Boeloni et la.…”

Section: Discussionunclassified

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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“…5,6 The use of autologous and allogeneic MSCs led to increased healing of critical sized bone defects. 7,8 MSCs implants enhanced the repair of experimentally …”

Section: Introductionmentioning

confidence: 99%

“…In one report, a dog with a nonunion bone defect received a percutaneous implant of autologous BM-MSCs, and after 8 weeks the patient was able to use its leg. 10 MSCs exhibit three characteristics -angiogenesis, 11 multipotentiality, 5,12 and immunomodulation 9,13 -that makes them very attractive and suitable for tissue engineering purposes. Those cells could be used as an alternative to traditional treatment in veterinary medicine.…”

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confidence: 99%

Allogeneic stem cell transplantation for bone regeneration of a nonunion defect in a canine

Yaneselli¹,

Filomeno²,

Semiglia³

et al. 2013

VMRR

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Nonunion bone defects occur frequently with local pain, functional limitations, muscular atrophy, and fistulas due to osteomyelitis. The application of mesenchymal stem cells (MSCs) could improve regeneration of bone following bone defects. The objective of the present study was to evaluate the treatment of a nonunion defect due to chronic osteomyelitis in a greyhound female dog with allogeneic adipose tissue-derived mesenchymal stem cells (AT-MSCs). The implanted cells were adherent to plastic, were of fibroblast type, and expressed the canine stem cell markers CD90 low , CD44 high , and CD45 -. Cell therapy consisted of five percutaneous weekly injections of 2 × 10 6 allogeneic AT-MSCs into the bone defect (total of 10 × 10 6 AT-MSCs).The patient was evaluated clinically and radiologically for up to 1 year. The results were clinical improvement, a light lameness score of 1 at week 16, return to use of its forearm, no pain, and increased muscular mass. No signs of osteomyelitis were observed radiologically and clinically there were no fistulas. There was no evidence of local or systemic adverse reactions caused by the alloimplants. The clinical relevance of the cell therapy contributing to repair of bone defects in small animals is a very promising future alternative. These results may have an important impact in new regenerative treatments for animal and human orthopedics.

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Canine adipose-derived-mesenchymal stem cells do not lose stem features after a long-term cryopreservation

Cited by 122 publications

References 23 publications

Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

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

Allogeneic stem cell transplantation for bone regeneration of a nonunion defect in a canine

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