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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Development of pipette tip gap closure migration assay (s-ARU method) for studying semi-adherent cell lines

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Author(s):
Sanmukh, Swapnil Ganesh [1] ; Felisbino, Sergio Luis [1]
Total Authors: 2
Affiliation:
[1] Univ Estadual Paulista Julio de Mesquita Filho UN, Inst Biosci Botucatu, Dept Morphol, Rua Antonio Celso Wagner Zanin 250, BR-18618689 Botucatu, SP - Brazil
Total Affiliations: 1
Document type: Journal article
Source: Cytotechnology; v. 70, n. 6, p. 1685-1695, DEC 2018.
Web of Science Citations: 0
Abstract

This work presents a pipette tip gap closure migration assay prototype tool (semi-adherent relative upsurges-ARUmethod) to study cell migration or wound healing in semi-adherent cell lines, such as lymph node carcinoma of the prostate (LNCaP). Basically, it consists of a 6-well cover plate modification, where pipette tips with the filter are shortened and fixed vertically to the inner surface of the cover plate, with their heights adjusted to touch the bottom of the well center. This provides a barrier for the inoculated cells to grow on, creating a cell-free gap. Sucha uniform gap formed can be used to study migration assay for both adherent as well as semi-adherent cells. After performing time studies, effective measurement of gap area can be carried out conveniently through image analysis software. Here, the prototype was tested for LNCaP cells, treated with testosterone and flutamide as well as with bacteriophages T4 and M13. A scratch assay using PC3 adherent cells was also performed for comparison. It was observed that s-ARU method is suitable for studying LNCaP cells migration assay, as observed from our results with testosterone, flutamide, and bacteriophages (T4 and M13). Our method is a low-cost handmade prototype, which can be an alternative to the other migration assay protocol(s) for both adherent and semi-adherent cell cultures in oncological research along with other biological research applications. (AU)

FAPESP's process: 16/09532-3 - Prostate cancer: changes in the Syndecan proteoglycans, oxidative stress enzyme sulfiredoxin and cell cycle kinase MELK
Grantee:Sérgio Luis Felisbino
Support Opportunities: Regular Research Grants