[Modifications in cultivating gingival keratinocytes]

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.
• 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.