• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Master's Dissertation
DOI
https://doi.org/10.11606/D.85.2011.tde-01062011-153618
Document
Author
Full name
Daniele Yoshito
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Mathor, Monica Beatriz (President)
Isaac, Cesar
Rodas, Andrea Cecilia Dorion
 
Title in Portuguese
Cultivo e irradiação de fibroblastos humanos em meio enriquecido com lisado de plaquetas para obtenção de camada de sustentação em cultura de células de epiderme
Keywords in Portuguese
cultura celular
feeder layer
fibroblastos humanos
lisado de plaquetas
Abstract in Portuguese
Por mais de 30 anos, a utilização do meio de cultura, enriquecido com soro bovino, e de fibroblastos murinos, com a taxa de proliferação controlada por irradiação ou por ação de drogas anti-cancerígenas, vem desempenhando com sucesso o seu papel de auxiliar no desenvolvimento dos queratinócitos em cultura, para fins clínicos. Porém, atualmente há uma preocupação crescente acerca da possibilidade de transmissão de príons e virose animais, aos pacientes transplantados. Levando em conta esta preocupação, o presente trabalho tem como objetivo cultivar fibroblastos humanos em meio enriquecido com lisado de plaquetas humanas e determinar a dose de irradiação dessas células, para obtenção da camada de sustentação na cultura de células da epiderme. Para realização do objetivo proposto, padronizamos a lise das plaquetas, utilizamos este lisado para cultivar os fibroblastos humanos e verificamos a dose de irradiação suficiente para inibir sua duplicação. Queratinócitos humanos foram cultivados nestas camadas de sustentação, em meio de cultura suplementado com o lisado. Com os resultados obtidos concluímos que o lisado de plaquetas a 10% promoveu uma melhor adesão e proliferação dos fibroblastos humanos e em todas as doses testadas (60 a 300 Gy), estes tiveram as suas atividades mitóticas inativadas pela radiação ionizante, sendo que as camadas de sustentação obtidas com doses de 70 a 150 Gy foram as que proporcionaram o melhor desenvolvimento dos queratinócitos em meio contendo 2,5% de lisado de plaquetas humanas. Portanto, foi possível padronizar, tanto o cultivo dos fibroblastos humanos, quanto sua inativação para utilização como camada de sustentação na cultura de queratinócitos, de maneira a eliminar os componentes xenobióticos.
 
Title in English
Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture
Keywords in English
cell culture
feeder layer
Abstract in English
For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
Publishing Date
2011-06-21
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors.
CeTI-SC/STI
© 2001-2024. Digital Library of Theses and Dissertations of USP.