Extraction and identification of primary macrophages from human hepatocellular carcinoma

Objective To study the extraction and identification of primary tumor-associated macrophages (TAMs) from human hepatocellular carcinoma (HCC). Methods Fifteen hepatocellular carcinoma specimens were collected from patients with primary HCC. Immunohistochemical staining was used to detect the expression of tumor-related macrophage surface markers CD68 and CD206 in HCC tissues. TAMs were separated by mechanical method and enzymatic digestion method. After 24 hours of adherent culture, the cell morphology was observed under inverted microscope. CD68 and CD206 were used as markers to identify the purity of TAMs by flow cytometry. Results CD68 and CD206 were positive in the central and adjacent tissues, and the positive cells were brown. A certain purity primary macrophages were obtained with morphological characteristics of macrophages. The single positive rate of CD68 was 36.29%, and the double positive rate of CD68 and CD206 was 15.96%. Conclusion The method used in this study can effectively extract and cultivate primary TAMs from human HCC tissues, which is of great value to further study the characteristics of TAMs and their effects on biological behavior of tumors. Key words: Macrophages; Carcinoma, hepatocellular; Cell culture techniques; Extraction; Identification