Hi researchers,
I have 2 questions about growing SHSY5Y cells:
1. I am currently growing SHSY5Y cells in culture and just wanted to check with those who work on this cell line. Is the morphology of my cells fine (please refer to attached images)? These are undifferentiated cells and should exhibit more clustering than if they were differentiated with Retinoic acid. However, I still do see neuritic extensions in addition to the epithelial morphology. So I am not sure if the cells are growing as they should be.
I currently maintain the cells with complete DMEM (1X DMEM with glucose, 10% FBS and 1% PenStrep). I usually freeze with 70% complete DMEM, 20% FBS and 10% DMSO. I always perform a cell count before splitting and the viabilities have been above 93%.
2. In addition, sometimes when I do not need the cells for an experiment in the immediate week, I typically seed a very small cell number (~400,000 cells in T75 flask), just changing media once every 3 days. But by the end of one week, I usually find the cell number depleting and media turning cloudy. From what I had researched, it is said that this cell line should never be seeded at such low densities and media should be changed frequently. Is this accurate?
I have been now seeding at higher densities and I don't encounter the issue of the media turning cloudy.
Thank you!!
It appears healthy and the morphology seems fine. You could always refer to ATCC and other investigators' previously reported images for such details.
Hello Mathangi Palanivel
1. The morphology of SHSY5Y cells is fine. In the undifferentiated form, SHSY5Y cells are characterized morphologically by neuroblast-like, non-polarized cell bodies with few, truncated processes. The cells tend to grow in clusters and may form clumps as cells appear to grow on top of one another in the central region of a cell mass. At edges of the cluster, cells begin to extend short neurites.
2. What you have researched is accurate. You will need to plate SHSY5Y cells at a density conducive to cell–cell communication to proliferate. If cells are plated too sparsely, growth rate is reduced, and cell death will be high.
Best.
Thomas Gantert
Thank you so much for the elaborate answer! I did recently encounter a loss of cells when I plated at a low density, but what I am growing currently is thawed from a new batch and I would suppose independent from that which experienced the loss.
Malcolm Nobre Ok thank you! Good to know:) Will bear in mind about the seeding density and media changes for this cell line. On a side note do you have experience differentiating SHSY5Y cells using retinoid acid? (or perhaps Thomas Gantert
Thomas Gantert
Thank you for the insights! Would it be possible to share your TPA-based differentiation protocol so that I can try it? Thank you!!- Badges
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