Proliferating Fibroblasts and HeLa Cells Co-cultured In Vitro Reciprocally Influence Growth Patterns, Protein Expression, Chromatin Features and Cell Survival

摘要: Aim: to identify biological interactions between proliferating fibroblasts and HeLa cells in vitro. Materials Methods: Fibroblasts were isolated from both normal tumour human tissues. Coverslip co-cultures of various ratios with medium replacement every 48 h studied using fixed cell staining dyes such as Giemsa silver staining, immunochemistry for Ki-67 E-cadherin, dihydrofolate reductase (DHFR) enzyme reaction, well live non-specific esterases lipids. Other techniques included carmine labeling, autoradiography apoptosis assessment. Results: Under conditions feeding cell: allowing parallel growth cells, co-cultured up 20 days, a series phenomena occur consecutively: profound affinity the two types exchange small molecules; encircling colonies by enhanced at their contact areas; expression carbonic anhydrase high cytoplasmic argyrophilia surrounding fibroblasts; intense production secretion lipid droplets development complex net argyrophilic projections E-cadherin cells; 10th day onwards, an increasing detachment batches peripheries appearance areas many multi-nucleated apoptotic fragments; 17th day, blocked G2-M phase. Co-cultures approximately 17-20 days display cell-cell fight foci (a) sparse types, (b) overgrowth (c) regrowth colonies. These results indicate that during interaction vitro, can be activated against HeLa. This type activation is not observed if fibroblast proliferation inhibition confluency, or omitting nutrient medium. Conclusion: The present observations show that: vitro drastically influences each other's protein expression, pattern, chromatin features survival; these functions depend on fibroblast/HeLa ratio, topology (cell-cell architectural pattern developed co-culture) frequent change, prerequisites proliferation; this co-culture model useful study processes within microenvironment, reproduction several conventionally seen cytological sections, desmoplasia, apoptosis, nuclear abnormalities; (d) overgrown adhering boundaries produce secrete droplets.