Publicação
Osteogenic differentiation of human umbilical cord-derived stromal cells on human amniotic membrane
| Resumo: | Mesenchymal stromal cells (MSC) from the umbilical cord (UC) are a promising tool for regenerative medicine due to their easy acquisition, high expansion potential and the ability to differentiate into multiple lineages. In particular, the capacity for osteogenic differentiation makes them of great interest for clinical application in bone repair. In this work, 51 MSC lines were isolated from different parts of 26 UC and successfully expanded in fetal bovine serum (FBS)-containing medium. These cells complied with the criteria for MSC classification as they were adherent to plastic, capable of multilineage differentiation, and expressed CD73, CD90, CD44 and CD105 while lacking hematopoietc and endothelial markers. Differentiation of the different MSC lines along the osteogenic lineage was variable depending mostly on the tissue source and isolation method. Moreover, osteogenic differentiation could be modulated by selecting for cells with high expression levels of CD105 (Endoglin) or by the co-culture of MSCs with different osteogenic potential. With the aim of establishing xeno-free clinically compatible culture conditions, StemPro MSCs serum free medium (SP-MSC SFM), human platelet lysate (in medium or in plasma), human plasma and AB serum (hABS) were tested. We have observed that only hABS supplemented medium could efficiently sustain MSC expansion, morphology, immunophenotype and multi-lineage differentiation similarly to FBS-containing medium. Finally, preliminary data indicates that frozen immunologically inert amniotic membrane is a suitable substrate for osteogenic differentiation of MSCs and, therefore, a promising scaffold for producing an osseous matrix for application in regenerative medicine. |
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| Autores principais: | Oliveira, Ana Rita Almeida, 1988- |
| Assunto: | Diferenciação celular Membrana amniótica Medicina regenerativa Cordão umbilical Teses de mestrado - 2011 |
| Ano: | 2011 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Mesenchymal stromal cells (MSC) from the umbilical cord (UC) are a promising tool for regenerative medicine due to their easy acquisition, high expansion potential and the ability to differentiate into multiple lineages. In particular, the capacity for osteogenic differentiation makes them of great interest for clinical application in bone repair. In this work, 51 MSC lines were isolated from different parts of 26 UC and successfully expanded in fetal bovine serum (FBS)-containing medium. These cells complied with the criteria for MSC classification as they were adherent to plastic, capable of multilineage differentiation, and expressed CD73, CD90, CD44 and CD105 while lacking hematopoietc and endothelial markers. Differentiation of the different MSC lines along the osteogenic lineage was variable depending mostly on the tissue source and isolation method. Moreover, osteogenic differentiation could be modulated by selecting for cells with high expression levels of CD105 (Endoglin) or by the co-culture of MSCs with different osteogenic potential. With the aim of establishing xeno-free clinically compatible culture conditions, StemPro MSCs serum free medium (SP-MSC SFM), human platelet lysate (in medium or in plasma), human plasma and AB serum (hABS) were tested. We have observed that only hABS supplemented medium could efficiently sustain MSC expansion, morphology, immunophenotype and multi-lineage differentiation similarly to FBS-containing medium. Finally, preliminary data indicates that frozen immunologically inert amniotic membrane is a suitable substrate for osteogenic differentiation of MSCs and, therefore, a promising scaffold for producing an osseous matrix for application in regenerative medicine. |
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