A method for establishing human primary gastric epithelial cell culture from fresh surgical gastric tissues.
At present, biopsy specimens, cancer cell lines and tissues obtained by gastric surgery are used in the study and analysis of gastric cancer, including the molecular mechanisms and proteomics. However, fibroblasts and other tissue components may interfere with these techniques. Therefore, the present study aimed to develop a procedure for the isolation of viable human gastric epithelial cells from gastric surgical tissues. A method was developed to culture human gastric epithelial cells using fresh, surgically excised tissues and was evaluated using immunocytochemistry, periodic acid-Schiff (PAS) staining and cell viability assays. Low cell growth was observed surrounding the gastric tissue on the seventh day of tissue explant culture. Cell growth subsequently increased, and at 12 days post-explant a high number of pure epithelial cells were detected. The gastric cancer cells exhibited rapid growth with a doubling time of 13-52 h, as compared to normal cells, which had a doubling time of 20-53 h. Immunocytochemical analyses of primary gastric cells revealed positive staining for cytokeratin 18 and 19, which indicated that the culture was comprised of pure epithelial cells and contained no fibroblasts. Furthermore, PAS staining demonstrated that the cultured gastric cells produced neutral mucin. Granulin and carbohydrate antigen 724 staining confirmed the purity of gastric cancer and normal cells in culture. This method of cell culture indicated that the gastric cells in primary culture consisted of mucin-secreting gastric epithelial cells, which may be useful for the study of gastric infection with Helicobacter pylori and gastric cancer.
Department of Biochemistry and Molecular Biology, Dalian Medical University, Liaoning Provincial Core Lab of Glycobiology and Glycoengineering, Dalian, Liaoning 116044, P.R. China.
Faisal
F
Aziz
Department of Biochemistry and Molecular Biology, Dalian Medical University, Liaoning Provincial Core Lab of Glycobiology and Glycoengineering, Dalian, Liaoning 116044, P.R. China.
Xuesong
X
Yang
Department of Anesthesia, The First Affiliated Hospital, Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
Qingping
Q
Wen
Department of Biochemistry and Molecular Biology, Dalian Medical University, Liaoning Provincial Core Lab of Glycobiology and Glycoengineering, Dalian, Liaoning 116044, P.R. China.
Qiu
Q
Yan
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism; D002478:Cells, Cultured; D005753:Gastric Mucosa / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D053538:Keratin-18 / Q000378:metabolism; D053539:Keratin-19 / Q000378:metabolism; D008875:Middle Aged; D009077:Mucins / Q000378:metabolism; D061251:Primary Cell Culture / Q000379:methods*; D000077153:Progranulins; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D013270:Stomach / Q000166:cytology*; D013274:Stomach Neoplasms / Q000378:metabolism / Q000473:pathology
D015295:Antigens, Tumor-Associated, Carbohydrate; C083825:CA-72-4 antigen; D036341:Intercellular Signaling Peptides and Proteins; D053538:Keratin-18; D053539:Keratin-19; D009077:Mucins; D000077153:Progranulins; D018809:Proliferating Cell Nuclear Antigen