Therefore, fresh gastric surgical tissue from patients of lower age was recommended to produce cultures of high growth rate and with low risk of contamination. Cell adherence to the flask surface is another important factor for the establishment of a successful primary culture 8. Fibronectin increases the rate of attachment and growth of the gastric epithelial cells. A previous study reported that fibronectin was a suitable substrate for mediating cell attachment 8. During primary gastric culture, there are frequently problems with gastric stromal tissue components, including fibroblasts, which have potential cytotoxic effects and perturb the growth of pure gastric epithelial cells In the present study, when cells surrounding the tissue began to appear following seven days of culture, no fibroblast cells were observed as the tissue was cultured in starting media, which contained no serum.
Fibroblast growth depends upon the presence of serum; however, this is not a requirement for the growth of gastric cancer cells For this reason, only gastric epithelial cell growth rather than fibroblast growth was observed in the cultured tissues. At a later stage, conventional media was used to promote rapid cell growth, and concurrent slow growth of fibroblast cells was detected 7. The culture medium may modulate the biological behavior of cultured cells The appropriate use of media is an important factor for the successful growth and differentiation of gastric epithelial cells 8.
It is also recognized that the majority of cells in vivo secrete endogenous growth factors to stimulate their own proliferation 8. Therefore, similar conditions may be achieved by using the appropriate media conditions during cell culture. In the present study, RPMI media, which is characterized by a combination of richness in trace elements, amino acids and high nutrient concentration, was used. Sodium bicarbonate and hepes buffer were also used for their abilities to maintain the pH of the media The characterization and investigation of cytokeratin expression by gastric cells is a simple method of determining epithelial nature 5.
Cytokeratin markers were therefore used in the present study, to confirm that the primary gastric cells in culture were free from fibroblasts and comprised a pure epithelial gastric cell population. High levels of staining for cytokeratin 18 and 19 were identified in gastric normal and cancer primary cells, which confirmed the epithelial nature of the gastric culture cells.
The cells that did not stain with cytokeratin 18 or 19 were presumed to be fibroblast cells 5. Mucin, which is found within the cells, may also be used to characterize gastric primary cultures 9. The PAS staining method was used to determine mucin expression within the gastric cells. In the present study, purple cytoplasmic staining was detected, which indicated the presence of neutral mucin within the gastric epithelial cells. This combination of neutral mucin and cytokeratin 18 and 19 expression demonstrated that the primary cultures were comprised of mucin-secreting gastric epithelial cells 5.
In order to differentiate gastric cancer cells from gastric normal cells, the expression of CA and GRN, which are associated with gastric tumor cells, was evaluated. High levels of CA and GRN staining are detected in gastric cancer cells, compared to those in normal gastric cells CA is a specific gastric cancer marker used for the diagnosis of gastric diseases Chen et al 18 demonstrated that CA was the most correlative and specific tumor biomarker for gastric cancer in the Chinese population. GRN is also a gastric cancer marker, which is highly expressed in gastric cancer and promotes cell proliferation, migration and invasion Determination of the gastric cell proliferation rate aids the elucidation of the replicative ability of the primary cell culture.
In order to determine the gastric culture cell proliferation rate, immunocytochemical staining was performed using PCNA antibodies. The results showed that difference in replication rate between gastric cancer and normal cells lies in the S-phase progression of the cell cycle. The replication rate of gastric cancer cells was higher than that of normal gastric cells. These results indicated that primary gastric cells had an active DNA synthesis and possessed the potential to continue gastric epithelial cell replication.
Cell growth was examined using a trypan blue exclusion assay. Gastric cancer cell growth was markedly higher 13—52 h than that of normal gastric cells 20—53 h. This result indicated that gastric cancer cells grow more rapidly than normal gastric cells 5. In conclusion, the present study provided a method for the primary culture of gastric epithelial cells from fresh gastric surgical tissue.
The advantage of the gastric primary culture method outlined is that the human tissue remained in medium and kept its activity intact with sufficient nutrition and adherence to the flask, which provided a suitable environment for continuous cell growth. The growth rate of gastric epithelial cells using this protocol was high and cultures were free from fibroblast cells.
These cultured gastric epithelial cells may therefore be used to investigate the effects of H. The present study was supported by the China grant no. Aziz F, Sherwani SK, Akhtar SS and Kazmi SU: Development of an in-house enzyme-linked immunosorbent assay based on surface whole cell antigen for diagnosis of Helicobacter pylori infection in patients with gastroduodenal ulcer disease.
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Isolation and Culture of Human Endometrial Epithelial Cells
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Robust cell growth for your experiments
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Mini Review ARTICLE
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SuperCult® Complete Mouse Epithelial Cell culture Medium Kit
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- Tissue Culture of Epithelial Cells | Mary Taub | Springer.
Stem Cells Transl Med 3 , —, doi: Wang, Z. PLoS One 11 , e, doi: We have a dedicated site for Germany. Epithelial cells are present in many different tissues in the body, and possess a diverse number of functional properties. However, all epithelial cells share some common characteristics. The cells possess a morphological polarity an-apical and basolateral surface , and are interconnected by tight junctions. The epithelial cells also possess the capacity to transport select solutes across the monolayer.
Such characteristics of epithelial cells can be examined in the tissue culture situation. This volume discusses the use of cell culture techniques to study these fundamental properties of epithelial cells.