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[Modifications in cultivating gingival keratinocytes]

G Lauer1, J E Otten, W Schilli

  • 1Klinik und Poliklinik für Mund-, Kiefer- und Gesichtschirurgie, Albert-Ludwigs-Universität, Freiburg.

Mund-, Kiefer- Und Gesichtschirurgie : MKG
|February 1, 1997
PubMed
Summary

Researchers compared gingival keratinocyte culture methods, finding explant cultures superior to single-cell suspensions. Autogenous serum proved as effective as fetal calf serum for keratinocyte growth and morphology.

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Area of Science:

  • Oral biology
  • Tissue engineering
  • Regenerative medicine

Background:

  • Current gingival keratinocyte culture methods utilize xenogenic materials like mouse fibroblasts and fetal calf serum.
  • These xenogenic additives pose risks, including potential DNA and protein contamination.
  • Safer alternatives are needed for culturing gingival autografts and for specialized cell cultures.

Purpose of the Study:

  • To compare the explant culture technique with the disperse culture technique for gingival keratinocytes.
  • To evaluate the efficacy of autogenous human serum as a substitute for fetal calf serum in keratinocyte culture.
  • To assess the growth, morphology, and cell biological characteristics of gingival keratinocytes under different culture conditions.

Main Methods:

  • Gingival biopsies were divided; one half was processed into single-cell suspensions, the other as explants.

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  • Cultures were established using either autogenous human serum or fetal calf serum.
  • Growth, epithelial layer confluency, morphology, and cytokeratin expression were analyzed.

    • Main Results:

    • Explant cultures consistently formed confluent primary gingival keratinocyte layers.
    • Single-cell suspensions failed to produce confluent epithelial layers.
    • Gingival keratinocyte growth and morphology were equivalent in autogenous serum and fetal calf serum.
    • Cytokeratin expression remained identical across tested conditions.

    Conclusions:

    • The explant technique is superior for establishing confluent primary gingival keratinocyte cultures compared to single-cell suspensions.
    • Autogenous human serum supports gingival keratinocyte growth and maintains normal morphology and cytokeratin expression, comparable to fetal calf serum.
    • Combining the explant technique with autogenous serum offers a safe and effective method for culturing gingival autografts and for specialized applications.