Monolayer formation and DNA synthesis of the outer epithelial cells from pearl oyster mantle in coculture with amebocytes

Masahiko Awaji, Tohru Suzuki

Research output: Contribution to journalArticlepeer-review

32 Citations (Scopus)

Abstract

In vitro experiments were conducted to clarify the involvement of the epithelium-amebocyte interaction in epithelial i regeneration of bivalves. The outer epithelia of the pallial mantle of the pearl oyster, Pinctada fucata martensii, were separated in cell sheets from the inner connective tissue layers by digestion with Dispase. Clumps of the separated mantle epithelia were inoculated onto the amebocyte layers prepared on the bottom of culture dishes and maintained at 20°C in 5% CO2:95% air for 1 wk. Balanced salt solution with 0.03% (wt/vol) glucose was used as a culture medium. The epithelial cells adhered to the amebocyte layers within 24 h, changed their shape from cuboidal to squamous, and migrated and formed monolayer sheets within 3 d. Electron microscopy confirmed maintenance of epithelial polarity and cell to cell junction in the sheets; 6 d after the inoculation, 5-bromo- 2'-deoxyuridine was added to the culture at 30 μM. After labeling for 24 h, the cultures were fixed and stained with anti 5-bromo-2'-deoxyuridine antibody. Cells with immunoreactive nuclei were clearly observed in the epithelial cell sheets, indicating active DNA synthesis in the epithelial sheets. Thus, cocultured with amebocytes, the outer epithelial cells from pallial mantle tissue formed a monolayer sheet and started DNA synthesis. The morphological features of the mantle outer epithelial cells are analogous to those described for the in vivo cutaneous wound healing process, suggesting that the epithelium-amebocyte interaction is important in the regeneration of epithelium in bivalves.

Original languageEnglish
Pages (from-to)486-491
Number of pages6
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume34
Issue number6
DOIs
Publication statusPublished - 1998
Externally publishedYes

Keywords

  • Extracellular matrix
  • Growth factor
  • Mollusca
  • Wound healing

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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