0
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: not found

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

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          This work presents a pipette tip gap closure migration assay prototype tool (semi-adherent relative upsurge—s-ARU—method) 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. Such a 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.

          Related collections

          Author and article information

          Contributors
          + 55 14 3880 0474 , sergio.felisbino@unesp.br
          Journal
          Cytotechnology
          Cytotechnology
          Cytotechnology
          Springer Netherlands (Dordrecht )
          0920-9069
          1573-0778
          1 August 2018
          December 2018
          : 70
          : 6
          : 1685-1695
          Affiliations
          ISNI 0000 0001 2188 478X, GRID grid.410543.7, Department of Morphology, Institute of Biosciences of Botucatu, , Universidade Estadual Paulista “Júlio de Mesquita Filho” -UNESP, ; Rua Antonio Celso Wagner Zanin, 250, Botucatu, Sao Paulo 18618-689 Brazil
          Article
          PMC6269371 PMC6269371 6269371 245
          10.1007/s10616-018-0245-1
          6269371
          30069611
          © Springer Nature B.V. 2018
          Funding
          Funded by: FundRef http://dx.doi.org/10.13039/501100002322, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior;
          Award ID: 963-14-2
          Award Recipient :
          Funded by: FundRef http://dx.doi.org/10.13039/501100003593, Conselho Nacional de Desenvolvimento Científico e Tecnológico;
          Award ID: 305391/2014-3
          Award Recipient :
          Funded by: FundRef http://dx.doi.org/10.13039/501100001807, Fundação de Amparo à Pesquisa do Estado de São Paulo;
          Award ID: 2016/09532-3
          Award Recipient :
          Categories
          Method in Cell Science
          Custom metadata
          © Springer Nature B.V. 2018

          Comments

          Comment on this article