Fresh and cultured buccal cells as a source of mRNA and protein for molecular analysis

Michalczyk, Agnes, Varigos, George, Smith, Lance and Ackland, Leigh 2004, Fresh and cultured buccal cells as a source of mRNA and protein for molecular analysis, Biotechniques, vol. 37, no. 2, pp. 2-7.

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Title Fresh and cultured buccal cells as a source of mRNA and protein for molecular analysis
Author(s) Michalczyk, AgnesORCID iD for Michalczyk, Agnes
Varigos, George
Smith, Lance
Ackland, LeighORCID iD for Ackland, Leigh
Journal name Biotechniques
Volume number 37
Issue number 2
Start page 2
End page 7
Publisher Eaton Publishing
Place of publication Natick, Mass.
Publication date 2004-08
ISSN 0736-6205
Summary We developed a method for obtaining viable buccal cells from mouthwash samples for use as a source of mRNA and protein. Immunofluorescent analysis showed that most cells were derived from nonkeratinized parabasal epithelia, with a minor proportion of proliferative cells. Gene expression was detected in buccal cells using reverse transcription PCR, Western blot analysis, and immunofluorescence. Using a keratinocyte-specific medium, buccal cells could be cultured on Matrigel™-coated permeable filters for up to 2 weeks while maintaining the expression of some epithelial-specific markers, including cytokeratin 13, cytokeratin 10, transferrin receptor, and β-integrin. The basal marker cytokeratin 14 and Ki67, an indicator of cellular proliferation, were detected in a few cells. We show that buccal cells can be obtained from a noninvasive procedure for use as a source of material for biochemical analyses. A population of the buccal cells can be maintained in culture for up to 2 weeks using keratinocyte-specific medium in combination with extracellular matrix.
Language eng
Field of Research 060101 Analytical Biochemistry
Socio Economic Objective 970106 Expanding Knowledge in the Biological Sciences
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2004, Eaton Publishing
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