Dear Researchers,
I am reaching out to this esteemed community seeking insights into a recent experiment I conducted involving cell seeding in agarose micro-wells. My aim is to engage with fellow researchers who may shed light on the intriguing outcomes of my experiment.
In my recent experimentation, I seeded cells from the RD cell line into agarose micro-wells at a significantly high concentration. The resulting images closely resembled those depicted in published works. However, I found myself disheartened by the outcome as I had anticipated observing dense spheroids akin to those formed with fibroblasts – distinct, rounded structures with diameters smaller than the wells. Instead, the cells appeared densely packed within the micro-wells.
Upon reflection, I conjectured that the outcome I obtained merely represented physical cell packing under the influence of gravity within a confined space, lacking fidelity to in vivo conditions. However, my perspective shifted upon encountering a publication authored by members of this community. The depicted results closely mirrored my own, with cells spanning the entirety of the micro-wells, discernible to the naked eye. This instilled a sense of hope within me, suggesting that my results may not be as unsatisfactory as initially perceived. Nonetheless, doubts linger.
I am writing to seek your professional opinions on why such constructs are considered spheroids. Do they truly replicate in vivo conditions? Are they formed as a result of intercellular contacts, or are they merely conglomerations of cells within the micro-wells? Have you observed alterations in gene expression and surface protein profiles compared to monolayer cultures?
I believe your insights into these questions will greatly contribute to the advancement of our understanding in this field. Your expertise and guidance would be immensely appreciated.
Thank you for considering my inquiry. I eagerly anticipate your responses and the opportunity for fruitful discussion.
Warm regards. "#spheroid #agarose #microwell
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