Publicação
Exploiting hnMSC 3D cultures conditioned medium for tissue regeneration
| Resumo: | Wound healing is impaired in several chronic diseases, where several factors are responsible for non-healing wounds. Over the time, treatment for these disabling conditions remains limited and largely ineffective. Mesenchymal stromal cells are valid candidates for cell therapy due to their beneficial effects on tissue regeneration. Besides these effects that could be mediated by cellular proliferation and differentiation, the secretion of growth factors and cytokines that enhance tissue repair has gained more attention. UCX®, in particular, is a specific population of human neonatal umbilical cord matrix derived MSCs that was found promising in this field and which paracrine mechanisms, for promoting cutaneous healing, have been studied. In this work, UCX® 3D cultures were exploited for conditioned media (CM) production. In contrast to 2D cultures, 3D mimic the complex in vivo environment, thus CM obtained by this methodology is expected to be more efficient in promoting wound healing. CM was obtained from UCX® traditional monolayer (CM2D) and 3D cultures (CM3D) and concentrated up 10x. Cell viability/proliferation assays in keratinocytes (HaCaT) and dermal fibroblasts (HDF), were performed, proving that CM3D and CM2D 10x could be applied in the in vivo assays. In addition, the therapeutic potential of the CM was evaluated in vivo in a rat wound healing model. CM2D and CM3D were applied via subcutaneous injection, to dorsal full-thickness incisions. The wound closure was monitored daily by size measurements. Skin biopsies were also taken at several time points for histological analyses to assess and compare different healing stages. Results showed that CM-treated wounds presented healing enhancement when compared to controls. In particular, CM3D-treated wounds have an improvement of wound healing profile with regards to accelerated wound closure, re-epithelialization and granulation tissue formation. Overall, these results indicate that UCX®-derived CM from 3D cultures may have a place in novel therapies for skin regeneration. |
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| Autores principais: | Camões, Sérgio José Póvoas, 1991- |
| Assunto: | Células estaminais Biologia do desenvolvimento Diferenciação celular Regeneração dos tecidos Teses de mestrado - 2014 |
| Ano: | 2014 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso restrito |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | português |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Wound healing is impaired in several chronic diseases, where several factors are responsible for non-healing wounds. Over the time, treatment for these disabling conditions remains limited and largely ineffective. Mesenchymal stromal cells are valid candidates for cell therapy due to their beneficial effects on tissue regeneration. Besides these effects that could be mediated by cellular proliferation and differentiation, the secretion of growth factors and cytokines that enhance tissue repair has gained more attention. UCX®, in particular, is a specific population of human neonatal umbilical cord matrix derived MSCs that was found promising in this field and which paracrine mechanisms, for promoting cutaneous healing, have been studied. In this work, UCX® 3D cultures were exploited for conditioned media (CM) production. In contrast to 2D cultures, 3D mimic the complex in vivo environment, thus CM obtained by this methodology is expected to be more efficient in promoting wound healing. CM was obtained from UCX® traditional monolayer (CM2D) and 3D cultures (CM3D) and concentrated up 10x. Cell viability/proliferation assays in keratinocytes (HaCaT) and dermal fibroblasts (HDF), were performed, proving that CM3D and CM2D 10x could be applied in the in vivo assays. In addition, the therapeutic potential of the CM was evaluated in vivo in a rat wound healing model. CM2D and CM3D were applied via subcutaneous injection, to dorsal full-thickness incisions. The wound closure was monitored daily by size measurements. Skin biopsies were also taken at several time points for histological analyses to assess and compare different healing stages. Results showed that CM-treated wounds presented healing enhancement when compared to controls. In particular, CM3D-treated wounds have an improvement of wound healing profile with regards to accelerated wound closure, re-epithelialization and granulation tissue formation. Overall, these results indicate that UCX®-derived CM from 3D cultures may have a place in novel therapies for skin regeneration. |
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