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For primary antibodies and protocol details see Table 2. Either tissue explants or 1 × 105 cervical cells in passage one were grown on Superfrost Plus® slides (Gerhard Menzel, Glasbearbeitungswerk GmbH & Co. KG, Braunschweig, Germany) until confluence.
Already in passage one cells showed a decreased potential to form multilayers and extended passaging led to the loss of epitheloid cell character.
P0 primary cells, P1 cells in passage one The growth medium with the best results in the initial media testing was further modified.
However, the epithelial morphology was completely lost in passage 5. Already in passage one morphological alterations can be found in some parts of the monolayer.
Cell morphology of primary (five days after explantation) and cells in passage one (two days after passaging) was assessed by light microscopy.
Due to their spindle-shaped morphology in monolayer culture and because all explanted cells stained D7-FIB positive in passage one through passage four, we identified these cells as tendon-derived fibroblasts.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com