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1.
J Tissue Eng Regen Med ; 14(11): 1661-1672, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32893450

RESUMO

Prudent choices of cell sources and biomaterials, as well as meticulous cultivation of the tissue microenvironment, are essential to improving outcomes of tissue engineering treatments. With the goal of providing a high-quality alternative for bone and cartilage tissue engineering, we investigated the capability of bovine placental scaffolds to support adipose-derived cell differentiation into osteogenic and chondrogenic lineages. Decellularized bovine placenta, a high-quality scaffold with practical scalability, was chosen as the biomaterial due to its rich extracellular matrix, well-developed vasculature, high availability, low cost, and simplicity of collection. Adipose-derived cells were chosen as the cell source as they are easy to isolate, nontumorigenic, and flexibly differentiable. The bovine model was chosen for its advantages in translational medicine over the mouse model. When seeded onto the scaffolds, the isolated cells adhered to the scaffolds with cell projections, established cell-scaffold communication and proliferated while maintaining cell-cell communication. Throughout a 21-day culture period, osteogenically differentiated cells secreted mineralized matrix, and calcium deposits were observed throughout the scaffold. Under chondrogenic specific differentiation conditions, the cells modified their morphology from fibroblast-like to round cells and cartilage lacunas were observed as well as the deposit of cartilaginous matrix on the placental scaffolds. This experiment provides evidence, for the first time, that bovine placental scaffolds have the potential to support bovine mesenchymal stem cell adherence and differentiation into osteogenic and chondrogenic lineages. Therefore, the constructed material could be used for bone and cartilage tissue engineering.


Assuntos
Tecido Adiposo/citologia , Diferenciação Celular , Linhagem da Célula , Condrogênese , Osteogênese , Placenta/citologia , Alicerces Teciduais/química , Animais , Bovinos , Forma Celular , Feminino , Humanos , Células-Tronco Mesenquimais/citologia , Placenta/ultraestrutura , Gravidez
2.
Tissue Cell ; 50: 43-50, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29429517

RESUMO

Skin is an extensive and easily accessible organ possessing various cell types that are constantly renewed. Previous studies have suggested the presence of a stem cell niche at the bulge region of the hair follicle, which contains cells positive for CD200 and CD34. Thus, this study sought to identify these cell populations in canine skin cells using the following methods 1- collecting samples of adult and fetal skin and isolating and culturing these cells using a method of simple enzymatic digestion and 2- testing the cell cultures for CD200 and CD34 in vitro and comparing them with skin tissue samples (in situ). Immunofluorescence results were negative for both CD200 and CD34 in frozen and paraffin embedded tissue, whereas the analysis showed that cultured cells positive for CD34, CD200 and double positive cells could be visualized in different percentages. Additionally, the pluripotency marker OCT4 was positive in the isolated cells. Analysis of CD34, CD200 and OCT4 by RT-qPCR showed that there is expression in fetal and adult cells, although no difference was observed between groups. Our results suggest that bulge stem cells from both fetuses and adult dogs were reported with the use of CD34 and CD200 markers in this study, and further techniques for cell isolation and in vitro cultivation are needed in order to obtain enriched populations of skin stem cells in dogs.


Assuntos
Separação Celular/métodos , Folículo Piloso/citologia , Nicho de Células-Tronco/genética , Células-Tronco/citologia , Animais , Antígenos CD/genética , Antígenos CD34/genética , Linhagem da Célula/genética , Células Cultivadas , Cães , Regulação da Expressão Gênica no Desenvolvimento , Folículo Piloso/metabolismo , Queratinócitos/citologia , Fator 3 de Transcrição de Octâmero/genética , Células-Tronco/metabolismo
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