High-throughput production of tumor spheroids (melanoma and colon carcinoma) using simple plate treatment and automated fluorescence microscopy analysis

Página do item simplificado
dc.contributor.authorPRUDENTE, SULEYNA R.
dc.contributor.authorASSIS, JOAO V.A. de
dc.contributor.authorSANTOS, ESTHER C. dos
dc.contributor.authorSILVA, GIOVANA D. da
dc.contributor.authorRODRIGUES, ALEX A.
dc.contributor.authorROCHA, LEONARDO W.P. de S.
dc.contributor.authorFALCAO, PATRCIA L.
dc.contributor.authorVIEIRA, DANIEL P.
dc.coverageInternacional
dc.creator.eventoLATIN-AMERICAN CONGRESS OF ARTIFICIAL ORGANS AND BIOMATERIALS, 12th
dc.date.accessioned2024-03-14T19:02:34Z
dc.date.available2024-03-14T19:02:34Z
dc.date.eventoDecember 12-15, 2023
dc.description.abstractIntroduction and objective: Cancer is currently one of the leading causes of death in the world. The objective of this work is the formation of viable spheroids from cells of melanoma (SKMEL-37) and colon adenocarcinoma (HT29-MTX) cell lines and their evaluation regarding cell viability to enable the use of threedimensional cell culture as an alternative to the use of experimental animal models. Methodology: Cells were maintained in RPMI 1640 medium and kept in an incubator at 37°C, 5% CO₂ with controlled humidity. Upon reaching 60-70% confluence, cells were washed with phosphate buffered saline (PBS) and detached using trypsin solution. Afterwards, they were seeded in 24-well plates pre-treated with PluronicⓇ F-127 (0.5g/mL in 2-propanol) and turned back in incubator for 72 hours. Then, the formed spheroids were stained with Hoechst 33342 and SYTOX® Green solution, incubated for 60 minutes and images were acquired automatically in a HTS equipment (INCell 2500 HS, Cytiva). Results and discussion: Properly cohesive spheroids were obtained for both lineages, 20-30 per well. After 72h, only a small fraction of cells (about 5%) were considered unviable by SYTOX® staining. Principal Component Analysis (PCA) using 13 variables, and further Principal Component Regression (PCR) showed that nuclei mean and maximum intensities (Hoechst), and nuclei volume are the most relevant variables, corelated to number of plated cells. Days in culture appeared to not correlate with other variables. Conclusions: It was concluded that the methodology for the production of spheroids for melanoma and colon adenocarcinoma cell lines presented is simple, fast and cheap, in which, in 72 hours, the spheroids form freely, without restriction of shape and size and presenting low cell death, being also compatible with the high throughput screening technique (HTS). Nuclei volume and intensity can be used in future analysis to assess cell global viability in spheroids.
dc.event.siglaCOLAOB
dc.format.extent214-214
dc.identifier.citationPRUDENTE, SULEYNA R.; ASSIS, JOAO V.A. de; SANTOS, ESTHER C. dos; SILVA, GIOVANA D. da; RODRIGUES, ALEX A.; ROCHA, LEONARDO W.P. de S.; FALCAO, PATRCIA L.; VIEIRA, DANIEL P. High-throughput production of tumor spheroids (melanoma and colon carcinoma) using simple plate treatment and automated fluorescence microscopy analysis. In: LATIN-AMERICAN CONGRESS OF ARTIFICIAL ORGANS AND BIOMATERIALS, 12th, December 12-15, 2023, Mar del Plata, Argentina. <b>Abstract...</b> p. 214-214. Disponível em: https://repositorio.ipen.br/handle/123456789/47935.
dc.identifier.orcidhttps://orcid.org/0000-0002-0007-534X
dc.identifier.urihttps://repositorio.ipen.br/handle/123456789/47935
dc.local.eventoMar del Plata, Argentina
dc.rightsopenAccess
dc.titleHigh-throughput production of tumor spheroids (melanoma and colon carcinoma) using simple plate treatment and automated fluorescence microscopy analysis
dc.typeResumo de eventos científicos
dspace.entity.typePublication
ipen.autorJOAO VICTOR ALMEIDA DE ASSIS
ipen.autorESTHER CAROLINA DOS SANTOS
ipen.autorGIOVANA DIAS DA SILVA
ipen.autorALEX ALVES RODRIGUES
ipen.autorLEONARDO WILANS PEREIRA DE SOUZA ROCHA
ipen.autorPATRICIA LIMA FALCAO
ipen.autorDANIEL PEREZ VIEIRA
ipen.codigoautor15846
ipen.codigoautor15983
ipen.codigoautor15317
ipen.codigoautor15583
ipen.codigoautor15328
ipen.codigoautor16028
ipen.codigoautor3158
ipen.contributor.ipenauthorJOAO VICTOR ALMEIDA DE ASSIS
ipen.contributor.ipenauthorESTHER CAROLINA DOS SANTOS
ipen.contributor.ipenauthorGIOVANA DIAS DA SILVA
ipen.contributor.ipenauthorALEX ALVES RODRIGUES
ipen.contributor.ipenauthorLEONARDO WILANS PEREIRA DE SOUZA ROCHA
ipen.contributor.ipenauthorPATRICIA LIMA FALCAO
ipen.contributor.ipenauthorDANIEL PEREZ VIEIRA
ipen.event.datapadronizada2023
ipen.identifier.ipendoc30278
ipen.notas.internasAbstract
ipen.type.genreResumo
relation.isAuthorOfPublication0ef49894-e71c-48f1-be08-2f29de9b4c1b
relation.isAuthorOfPublication9d0ef255-9847-4920-91cc-51c240193f50
relation.isAuthorOfPublication5cd1e045-2814-47b2-b360-caf2443c0464
relation.isAuthorOfPublicationb93159e8-8c72-4ac5-bc5a-93911236b814
relation.isAuthorOfPublication30ea7198-caea-4d01-b34b-c57a11c2aa46
relation.isAuthorOfPublication009c64f6-ee20-44ce-a880-f52d8861c247
relation.isAuthorOfPublicationc2352608-be9c-4a73-be8c-571f10bb53d2
relation.isAuthorOfPublication.latestForDiscovery0ef49894-e71c-48f1-be08-2f29de9b4c1b
sigepi.autor.atividadeASSIS, JOAO V.A. de:15846:810:N
sigepi.autor.atividadeSANTOS, ESTHER C. dos:15983:-1:N
sigepi.autor.atividadeSILVA, GIOVANA D. da:15317:810:N
sigepi.autor.atividadeRODRIGUES, ALEX A.:15583:810:N
sigepi.autor.atividadeROCHA, LEONARDO W.P. de S.:15328:810:N
sigepi.autor.atividadeFALCAO, PATRCIA L.:16028:-1:N
sigepi.autor.atividadeVIEIRA, DANIEL P.:3158:810:N
Pacote Original
Agora exibindo 1 - 1 de 1
Imagem de Miniatura
Nome:
30278.pdf
Tamanho:
287.55 KB
Formato:
Adobe Portable Document Format
Baixar
Licença do Pacote
Agora exibindo 1 - 1 de 1
Nenhuma Miniatura disponível
Nome:
license.txt
Tamanho:
1.71 KB
Formato:
Item-specific license agreed upon to submission
Descrição:
Baixar
Coleções
Eventos - Resumos