Brazilian minipig as a large-animal model for basic research and stem cell-based tissue engineering. Characterization and in vitro differentiation of bone marrow-derived mesenchymal stem cells
J. appl. oral sci
;
22(3): 218-227, May-Jun/2014. tab, graf
Article
Dans Anglais
| LILACS, BBO
| ID: lil-711715
ABSTRACT
Stem cell-based regenerative medicine is one of the most intensively researched medical issues. Pre-clinical studies in a large-animal model, especially in swine or miniature pigs, are highly relevant to human applications. Mesenchymal stem cells (MSCs) have been isolated and expanded from different sources. Objective:
This study aimed at isolating and characterizing, for the first time, bone marrow-derived MSCs (BM-MSCs) from a Brazilian minipig (BR1). Also, this aimed to validate a new large-animal model for stem cell-based tissue engineering. Material andMethods:
Bone marrow (BM) was aspirated from the posterior iliac crest of twelve adult male BR1 under general anesthesia. MSCs were selected by plastic-adherence as originally described by Friedenstein. Cell morphology, surface marker expression, and cellular differentiation were examined. The immunophenotypic profile was determined by flow cytometry. The differentiation potential was assessed by cytological staining and by RT-PCR.Results:
MSCs were present in all minipig BM samples. These cells showed fibroblastic morphology and were positive for the surface markers CD90 (88.6%), CD29 (89.8%), CD44 (86.9%) and negative for CD34 (1.61%), CD45 (1.83%), CD14 (1.77%) and MHC-II (2.69%). MSCs were differentiated into adipocytes, osteoblasts, and chondroblasts as demonstrated by the presence of lipidic-rich vacuoles, the mineralized extracellular matrix, and the great presence of glycosaminoglycans, respectively. The higher gene expression of adipocyte fatty-acid binding protein (AP2), alkaline phosphatase (ALP) and collagen type 2 (COLII) also confirmed the trilineage differentiation (p<0.001, p<0.001, p=0.031; respectively).Conclusions:
The isolation, cultivation, and differentiation of BM-MSCs from BR1 makes this animal eligible as a useful large-animal model for stem cell-based studies in Brazil. .
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Porc miniature
/
Cellules de la moelle osseuse
/
Modèles animaux
/
Ingénierie tissulaire
/
Cellules souches mésenchymateuses
Type d'étude:
Étude pronostique
Limites du sujet:
Animaux
Pays comme sujet:
Amérique du Sud
/
Brésil
langue:
Anglais
Texte intégral:
J. appl. oral sci
Thème du journal:
Dentisterie
Année:
2014
Type:
Article
Pays d'affiliation:
Brésil
Institution/Pays d'affiliation:
University of São Paulo/BR
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