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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2014.tde-09122014-080415
Document
Author
Full name
Natalia Langenfeld Fuoco
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Paes, João Tadeu Ribeiro (President)
Lizier, Nelson Foresto
Visintin, José Antonio
Title in Portuguese
Diferentes metodologias para isolamento, expansão e caracterização de células-tronco derivadas de tecido adiposo humano.
Keywords in Portuguese
Células-tronco
Colagenase
Digestão enzimática
Dissociação mecânica
Lisado de plaquetas
Soro fetal bovino (SFB)
Tecido adiposo
Abstract in Portuguese
Os procedimentos para uso clínico de células-tronco derivadas de tecido adiposo (CT-TA) exigem grandes quantidades de células, por isso, em geral os protocolos envolvem a expansão e cultura celular in vitro. No entanto, as metodologias utilizadas rotineiramente para o cultivo de CT-TA envolvem a utilização de componentes xenobióticos, como a colagenase e o soro fetal bovino (SFB), que representam riscos potencias de reações imunológicas e transmissão de doenças infecciosas. Sendo assim, pretendeu-se no presente estudo analisar diferentes parâmetros metodológicos para isolamento e expansão de CT-TA, na ausência de componentes xenobióticos. Para tanto, as células-tronco foram isoladas por digestão enzimática ou dissociação mecânica e submetidas à expansão na presença de SFB ou lisado de plaquetas humano (LP). Os resultados mostraram que a metodologia de dissociação mecânica representa uma alternativa viável e eficiente para cultivo de CT-TA, e que o emprego de LP como suplemento para o meio de cultura aumentou de forma significativa a proliferação celular. Em função desses resultados, pode-se concluir que é possível a implementação de técnicas de isolamento e expansão de CT-TA, prescindindo-se de componentes xenobióticos.
Title in English
Different methodologies for isolattion and cultivation human adipose-derived stem cells.
Keywords in English
Adipose tissue
Collagenase
Enzymatic digestion
Fetal bovine serum
Mechanical dissociation
Platelet lysate
Stem cell
Abstract in English
The procedures for the clinical use of adipose-derived stem cells (ASC) require large amounts of cells, so in general protocols involve culture and cell expansion in vitro.However, the methods routinely used for the culture of ASC involves the use of xenobiotic components, such as collagenase and fetal bovine serum (FBS), that may representing potential risk of immunological reactions and the risk of transmission of infectious diseases. Thus, it was intended in this study to analyze different methodological parameters for the isolation and expansion of ASC in the absence of xenobiotic components. For this, stem cells were isolated by enzymatic digestion and mechanical dissociation and were submitted to expansion in the presence of FBS or human platelet lysate (PL). The results showed that the mechanical dissociation method represents an effective alternative to growing ASC, and that the use of PL as a supplement to the culture medium significantly increased cellular proliferation. In view of these results, we can conclude that it is possible to implement techniques for isolation and expansion of ASC, dispensing xenobiotic components.
 
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Publishing Date
2014-12-11
 
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