Development of a New Approach to Aid in Visual Identification of Murine iPS Colonies Using a Fuzzy Logic Decision Support System

Autor(es)

Bassaneze, Vinicius ^[1] ; Sacramento, Chester Bittencourt ^[1] ; Freire, Rodolfo ^[2] ; De Alencar, Patricia Fernandes ^[1] ; Siqueira Ortega, Neli Regina ^[2] ; Krieger, Jose Eduardo ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Med, Lab Genet & Mol Cardiol LIM 13, Heart Inst InCor, Sao Paulo - Brazil

^[2] Univ Sao Paulo, Fac Med, Ctr Fuzzy Syst Hlth, Dept Pathol, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 8, n. 8 AUG 8 2013.
• Citações Web of Science: 1

Resumo

The a priori identification of induced pluripotent stem cells remains a challenge. Being able to quickly identify the most embryonic stem cell-similar induced pluripotent stem cells when validating results could help to reduce costs and save time. In this context, tools based on non-classic logic can be useful in creating aid-systems based on visual criteria. True colonies when viewed at 100x magnification have been found to have the following 3 characteristics: a high degree of border delineation, a more uniform texture, and the absence of a cracked texture. These visual criteria were used for fuzzy logic modeling. We investigated the possibility of predicting the presence of alkaline phosphatase activity, typical of true induced pluripotent stem cell colonies, after 25 individuals, with varying degrees of experience in working with murine iPS cells, categorized the images of 136 colonies based on visual criteria. Intriguingly, the performance evaluation by area under the ROC curve (16 individuals with satisfactory performance), Spearman correlation (all statistically significant), and Cohen's Kappa agreement analysis (all statistically significant) demonstrates that the discriminatory capacity of different evaluators are similar, even those who have never cultivated cells. Thus, we report on a new system to facilitate visual identification of murine-induced pluripotent stem cell colonies that can be useful for staff training and opens the possibility of exploring visual characteristics of induced pluripotent stem cell colonies with their functional peculiarities. The fuzzy model has been integrated as a web-based tool named ``2see-iPS{''} which is freely accessed at http://genetica.incor.usp.br/2seeips/. (AU)

• Processo FAPESP: 08/52436-9 - Reprogramação dirigida de células mesenquimais de tecido adiposo em células-tronco pluripotentes e cardiomiócitos
• Beneficiário: José Eduardo Krieger
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 08/52335-8 - Reprogramacao de fibroblastos em celulas-tronco pluripotentes e diferenciacao destas em cardiomiocitos por meio da expressao de fatores de transcricao cardiacos.
• Beneficiário: Chester Bittencourt Sacramento
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 08/52334-1 - Reprogramacao de celulas mesenquimais de tecido adiposo humano em celulas-tronco pluripotentes por meio de proteina de fusao tat.
• Beneficiário: Vinícius Bassaneze
• Modalidade de apoio: Bolsas no Brasil - Doutorado