Currently I am working with the neuroblastoma cell line SK-N-BE(2). Today I stained the nuclei using Hoechst and analyzed the cells under the microscope. I was surprised when I found that control cells (grown in RPMI with 10% FBS) appeared to have several nuclei.
I could not find any pictures showing Hoechst-stained SK-N-BE(2) cells but I found a publication from 1981 (Barnes et al.: The fine structure of continuous human neuroblastoma lines SK-N-SH, SK-N-BE(2), and SK-N-MC).
They wrote:
"In stationary phase SK-N-SH and SK-N-BE(2) cultures, some cells formed aggregates or clumps up to 12 cells deep […].Between the clumps the cells formed a monolayer as observed with the light microscope. However, electron micrographs showed that in such areas there was extensive overlapping of lateral cytoplasmic extensions of cells and many nuclei were sandwiched between flat extensions of several other cells; such areas are referred to as "pseudomonolayers." Individual cells in the pseudomonolayer were not identified before embedding and sectioning, and areas between clumps included both neuroblastlike and flat cells."
Has someone heard about pseudomonolayers before? Or has someone worked with SK-N-BE(2) and found similar results?
I have uploaded a word-file with Hoechst-stained SK-N-BE(2) cultured in RPMI+ 10% FBS.
Hi Roth,
It is not quiet unusual having polyneucleated cell mass/lumps for the neuroblastoma cell line SK-N-BE(2) in culture. Being cancerous like other cancer cells the cells under investigation looses its polarity and grows in multilayer in certain areas forming spheroids. This is why this type of cancerous cell lines are good model for 3D culture in hanging droplets.
Please see the attached ARVO abstract where this type of pseudomonolayer formation is reported in corneal epithelium culture.
Hope that helps.
Hei Amritlal Mandal,
thank yopu very much for your answer!
you write "It is not quiet unusual having polyneucleated cell mass/clumps for the neuroblastoma cell line SK-N-BE(2) in culture."
Thus, SK-N-BE(2) cells are not polynucleated but as they are growing as cell clumps (pseudomonolayer), the cells appear to be polynucleated.
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