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Master's Dissertation
DOI
https://doi.org/10.11606/D.17.2020.tde-19082020-093414
Document
Author
Full name
Rafaela Rossetti
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2020
Supervisor
Committee
Passos Junior, Geraldo Aleixo da Silva (President)
Freitas, Gileade Pereira
Oliveira, Carolina Caliári
Title in Portuguese
Caracterização in vivo de células-tronco mesenquimais murinas da medula óssea e do osso compacto quanto à capacidade de geração de nicho hematopoético heterotópico
Keywords in Portuguese
Células-tronco mesenquimais
Medula óssea
Nicho hematopoético heterotópico
Osso compacto
Abstract in Portuguese
O nicho hematopoético é o microambiente composto por diferentes tipos celulares e estímulos que regulam a autorrenovação, diferenciação e proliferação das células-tronco hematopoéticas. As células-tronco mesenquimais (CTMs) são importantes componentes do nicho hematopoético. As CTMs foram inicialmente estudadas a partir da medula óssea, sabe-se que são capazes de formar colônias quando cultivadas e de se diferenciarem dando origem às células dos tecidos adiposo, ósseo e cartilaginoso, bem como de originar nicho hematopoético heterotópico após implantadas. Essas células foram inicialmente descritas com localização perivascular na medula óssea, porém estudos posteriores demonstraram que as CTMs também estão presentes na região do endósteo. Então, foi sugerida a hipótese da existência de duas populações distintas de CTMs na medula óssea, sendo uma de localização endosteal, responsável principalmente pela geração dos tecidos esqueléticos durante a fase de desenvolvimento e pela homeostase do tecido e outra, de localização perivascular, que atuaria no reparo tecidual durante a fase adulta. Assim, nesse trabalho, CTMs isoladas da medula óssea (MO-CTMs) e do osso compacto (OCCTMs) de camundongos da linhagem C57BL/6GFP foram comparadas quanto às suas características in vitro e capacidade de originar um nicho hematopoético heterotópico. Foi observado que as CTMs apresentaram diferenças com relação à frequência de CFU-F, perfil imunofenotípico, capacidade de diferenciação e expressão gênica. Adicionalmente, a avaliação de três diferentes protocolos para a padronização do implante de CTMs, visando formação de nicho hematopoético, mostrou que o implante de Matrigel com grânulos de hidroxiapatita, células endoteliais e OC-CTMs foi capaz de proporcionar a geração de um nicho hematopoético heterotópico.
Title in English
In vivo characterization of murine mesenchymal stem cells from bone marrow and compact bone in relation to its heterotopic hematopoietic niche generation potential
Keywords in English
Bone marrow
Compact bone
Heterotopic hematopoietic niche
Mesenchymal stem cells
Abstract in English
The hematopoietic stem cell niche is the microenvironment composed of different types of cells and stimulus that regulate self-renewing, differentiation and proliferation of the hematopoietic stem cells. The mesenchymal stem cells (MSCs) are important components of the hematopoietic niche. The MSCs were previously studied from bone marrow, it's known that, in culture, they are able to form colonies and differentiate giving rise to cells of adipose, bone and cartilaginous tissue, as well as originate heterotopic hematopoietic niche when implanted. These cells were initially described with perivascular location in the bone marrow, although later studies have shown that the MSCs are also present in the endosteum region. So, it's been suggested the hypothesis of existing two distinct populations of MSCs in the bone marrow, one located in the endosteum region, being mainly responsible for generating bone tissues during the development phase and for tissue homeostasis, and another, of perivascular location, which would act in tissue repair during adulthood. Therefore, in this work, MSCs isolated from bone marrow (BM-MSCs) and compact bone (CBMSCs) from C57BL/6gfp mice were compared to their in vitro characteristics and their capacity to originate a heterotopic hematopoietic niche. It's been observed that the MSCs have shown differences related to their frequency of CFU-F, immunophenotypic profile, capacity of differentiation and gene expression. Additionally, the evaluation of three different protocols to standardize MSCs implantation, aiming the hematopoietic niche formation, has shown that implantation of Matrigel along with hydroxyapatite granules, endosteum cells and BM-MSCs were able to provide the generation of a heterotopic hematopoietic niche.
 
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RAFAELAROSSETTIco.pdf (6.11 Mbytes)
Publishing Date
2020-10-23
 
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