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Master's Dissertation
DOI
10.11606/D.10.2010.tde-01042011-154719
Document
Author
Full name
Dayane Alcântara
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Miglino, Maria Angélica (President)
Kfoury Junior, José Roberto
Maria, Durvanei Augusto
Title in Portuguese
Obtenção e caracterização de linhagem celular primária de osteossarcoma canino
Keywords in Portuguese
Metástases
Neoplasias
Osteossarcoma
Abstract in Portuguese
O osteossarcoma é um tumor ósseo maligno comum em cães, com preferência por raças de grande porte, tem alto potencial metastático e ocorre frequentemente no esqueleto apendicular. O diagnóstico é baseado na história clínica, exame físico, achados radiológicos e histopatológicos. O tratamento consiste na ressecção cirúrgica do tumor e o protocolo de tratamento de escolha é a amputação associada à quimioterapia. O osteossarcoma apresenta semelhanças em cães e humanos, portanto o osteossarcoma canino pode ser um modelo útil para estudar esta doença em humanos. O objetivo deste trabalho foi estabelecer e caracterizar a linhagem celular primária de osteossarcoma canino. Os fragmentos tumorais foram obtidos por meio de biópsias e excereses cirúrgicas do osteosarcoma, a confirmação diagnóstica realizada pelo exame histopatológico. Os fragmentos foram cultivados em meios de cultura DMEM-H, DMEM-Low, RPMI-1640 e MEM para obtenção das linhagens. Para análise da morfologia celular foi realizada a fotodocumentação das garrafas em microscopia invertida, microscopia eletrônica de transmissão e varredura. A caracterização dos marcadores de superfície, citoplasmáticos e nucleares, análise das fases do ciclo celular e potencial elétrico mitocondrial foram realizados por citometria de fluxo. Foram obtidos 6 tumores, sendo 5 osteossarcomas e um condrossarcoma, dos quais foram obtidas 3 linhagens OST-1, OST-3, e CDS1. Os meios de cultivo DMEM-H e MEM foram eficientes para obtenção e manutenção das linhagens. Morfologicamente as células OST-1, apresentaram aspecto fusiforme enquanto as células OST-3 apresentaram pleomorfismo celular. Os marcadores de superfície, citoplasmáticos e nucleares nas células de osteossarcoma canino OST-3 apresentaram-se diferencialmente expressos, como os marcadores de origem mesenquimal, o marcador de reparo de DNA, P53, não foi expresso como também é encontrado em inúmeras linhagens de osteossarcoma.
Title in English
Obtention and characterization of canine osteosarcoma primary cell line
Keywords in English
Metastasis
Neoplasms
Osteosarcoma
Abstract in English
Osteosarcoma is a common malignant bone tumor in dogs, with a preference for large breeds, has a high metastatic potential and occurs often in the appendicular skeleton. Diagnosis is based on clinical history, physical examination, radiological and histopatological findings. The treatment consists in surgical tumor resection, and treatment protocol choice is amputation associated with chemotherapy. Osteosarcoma has many similarities in dogs and humans, thus the canine osteosarcoma can be a usefull model to study this disease in humans. The aim of this work was to establish and characterize the canine osteosarcoma primary cell line. Tumor samples were obtained by surgical biopsy and the osteosarcoma confirmation was made by histopathological examination. Tumor fragments were cultured in DMEM-H, DMEM- LOW, RPMI-1640 and MEM culture media to establish the cell lines. Cell morphology analysis was carried out by photodocumentation in inverted microscopy and in transmission electron microscopy. The characterization of surface, cytoplasmic and nuclear markers and cell cycle phases, and mitochondrial electric potential analysis were performed by flow cytometry and analyzed with the Win MDI 2.8. Five osteosarcomas, 1 chondrosarcoma, and three cancer cell lines, OST-1, OST-3, and CDS1, were obtained. DMEM-H and MEM culture medium were efficient in cell lines establishing and maintenance. OST-1 cell line showed spindle shaped and OST-3 cell line showed pleomorphism. Surface, cytoplasmic and nuclear markers in the OST-3 canine osteosarcoma cell line were differentially expressed as mesenchimal origin markers, the DNA repair marker, P53, it was not expressed, it is also found in other osteossarcoma lines.
 
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Dayane_Alcantara.pdf (3.94 Mbytes)
Publishing Date
2012-10-05
 
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