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Reversible ultrastructural changes in human fibroblasts grown in hepes buffered MCDB-104 supplemented with human
Abstract:
Prolonged maintenance of human dermal fibroblasts in MCDB-104 medium supplemented with pooled human serum or platelet factor deficient preparations of human serum led to appearance of a large number of membrane bound inclusions, resembling lysosomes, and proliferation of small, Golgi associated vesicles. These inclusions did not appear if the cells were grown in Dulbecco-Vogt's modification of Eagle's minimal essential medium or in minimal essential medium supplemented with the same human serum fractions. Cells that acquired inclusions during a 10 d incubation in MCDB-104 subsequently lost these inclusions when transferred to Dulbecco'Vogt's medium for 4 d. Similar reversal of effects of MCDB-104 was also produced by MCDB-104 buffered with bicarbonate instead of HEPES.
Insights
Human dermal fibroblasts cultured in MCDB-104 medium with serum developed lysosome-like inclusions. These cellular changes were reversible and medium-dependent, highlighting the impact of specific cell culture conditions.
Area of Science:
- Cell Biology
- Tissue Culture
- Fibroblast Research
Background:
- Cell culture media composition significantly influences cellular morphology and function.
- Understanding fibroblast responses to different media is crucial for research applications.
Purpose of the Study:
- To investigate the ultrastructural changes in human dermal fibroblasts cultured in MCDB-104 medium.
- To determine the role of specific media components and buffering systems in cellular morphology.
Main Methods:
- Human dermal fibroblasts were cultured in MCDB-104 medium with human serum or serum fractions.
- Control cells were cultured in Eagle's minimal essential medium (MEM) or its modification.
- Cells were analyzed for the presence of intracellular inclusions and vesicles using electron microscopy.
Main Results:
- Prolonged culture in MCDB-104 with serum induced lysosome-like inclusions and Golgi-associated vesicles.
- These inclusions were absent in cells cultured in MEM or MEM with serum.
- Transferring cells from MCDB-104 to MEM reversed the formation of inclusions.
- Bicarbonate buffering of MCDB-104 also prevented inclusion formation.
Conclusions:
- The formation of lysosome-like inclusions in fibroblasts is dependent on the specific cell culture medium, particularly MCDB-104.
- Specific components or properties of MCDB-104, potentially related to buffering, induce these ultrastructural changes.
- These findings underscore the importance of careful selection of cell culture media for maintaining normal fibroblast morphology.