[Comparison of two methods for primary culture of epithelial cells from human bioptic tissue of nasopharyngeal carcinoma].

OBJECTIVE

To compare two methods for primary culture of the epithelial cells from human bioptic specimens of nasopharyngeal carcinoma (NPC), and preliminarily study the biological characteristics of the primarily cultured cells.

METHODS

Thirty-three bioptic specimens were collected from patients with pathologically confirmed NPC who received no previous radiotherapy or chemotherapy. Seventeen samples were cultured using tissue culture method, and tissue predigestion culture was used for the other 16 samples. The cells were cultured in Keratinocyte-SFM medium containing 2% fetal bovine serum. The two culture methods were compared for successful rate, attachment time and average time of cell growth. The biological features of the cultured cells were observed by reverse microscope, cytokeratin immunocytochemistry, growth curve and survival analysis.

RESULTS

The successful rate, attachment time and days for cell growth of the tissue culture method and tissue predigestion culture method were 23.5% (4/17) vs 62.5% (10/16), 4.47±0.48 h vs 7.88±1.01 h, and 13.75±1.5 days vs 8.3±4.55 days, respectively, showing significant differences between the two methods (P<0.05). The cells resulting from tissue predigestion method, with a average survival of 62.72 days, were characterized by a fusiform morphology with large nuclei, multiple nucleoli, and cytokeratins positivity. In contrast, the cells obtained by tissue culture method showed multilayer alignment without distinct cell structures and failed to grow consistently.

CONCLUSION

Compared with tissue culture method, tissue predigestion cell culture results in high successful rate and requires shorter cell growth time with longer attachment time. Keratinocyte-SFM medium supplemented with low concentrations of FBS is suitable for the growth of primarily cultured epithelial cells from NPC biopsy samples in vitro.