Related Experiment Videos
On disturbance of homeostasis in organ-cultured tissue
Abstract:
The intact membranous rat mesentery was cultured in Eagle's minimum essential medium containing no serum or only low concentrations of serum. The procedure is in some important respects superior to previous organ culture techniques. To estimate the extent of disturbance of homeostasis of the tissue in culture, the spontaneous mast-cell histamine release was quantitated after preculture preparation of the specimens and after different intervals in culture. Also, the proliferation of fibroblasts and mesothelial cells that predominate in the mesentery was assessed at 48 h by cytofluorometric quantitation of DNA in single-tissue cells. Spontaneous histamine release was time dependent during cultivation, amounting to ca. 50% at 48 h, and was affected by the medium used for moistening the tissue before cultivation. Culturing also brought about great spontaneous increase in the proliferation of fibroblasts and mesothelial cells, the rate being related to the concentration of serum. Addition of the mast-cell secretagogues 48/80 or polymyxin B at 1 h caused rapid release of 50 to 60% of the histamine and was followed by augmented proliferation in the serum-containing media. The spontaneous increase of cell proliferation in tissue culture may be causally related to mast-cell secretion. Further studies are needed to define factors influencing the spontaneous mast-cell secretion and the mast-cell-dependent mitogenesis in normal tissue cells.
Insights
Rat mesentery tissue culture demonstrated spontaneous mast cell histamine release and increased fibroblast and mesothelial cell proliferation. These changes were influenced by culture medium serum concentration and may be linked.
Area of Science:
- Tissue Engineering and Regenerative Medicine
- Cell Biology
- Histamine Signaling
Background:
- Organ culture techniques are crucial for studying tissue homeostasis and cellular responses in vitro.
- Mast cells play a significant role in inflammatory and homeostatic processes through histamine release.
- Fibroblast and mesothelial cell proliferation are key indicators of tissue health and response to culture conditions.
Purpose of the Study:
- To evaluate the homeostasis of rat mesentery tissue during organ culture.
- To quantify spontaneous mast cell histamine release and fibroblast/mesothelial cell proliferation in cultured mesentery.
- To investigate the impact of culture medium composition, specifically serum concentration, on these cellular events.
Main Methods:
- Intact rat mesentery was cultured in Eagle's minimum essential medium with varying serum concentrations.
- Spontaneous mast cell histamine release was measured over time using quantitative assays.
- Fibroblast and mesothelial cell proliferation was assessed at 48 hours via cytofluorometric DNA quantitation.
Main Results:
- Spontaneous histamine release from mast cells increased over time, reaching approximately 50% at 48 hours.
- Tissue culture induced a significant increase in fibroblast and mesothelial cell proliferation, dependent on serum concentration.
- Mast cell secretagogues (48/80, polymyxin B) induced rapid histamine release and augmented proliferation in serum-containing media.
Conclusions:
- Rat mesentery organ culture leads to time-dependent mast cell histamine release and increased cell proliferation.
- Serum concentration in the culture medium significantly influences both histamine release and cell proliferation.
- Mast cell secretion may be causally linked to the observed spontaneous increase in cell proliferation, warranting further investigation.