Hi, I'm culturing human bronchial smooth muscle cell (Lonza). Recently, I'm having problem with this cell when I seeded them in 96 well plates (BD brand). After serum starved for 1 day, I change the medium with my treatment drug dissolved in complete medium. However, my control group (with complete medium only) show a shrinkage of the cell. Attached is the photo that I took. Can anyone tell me what's the problem? Is it cell senescence? Or is the 96 well plate the problem?
Ps: Only certain wells have this problem. Some of the well (with complete medium only) have normal morphology in the same plate.
Huimin,
Do you think the cells are not attaching properly or dislodging from the bottom of the well and subsequently undergoing apoptosis?
I had trouble in the past culturing ASMCs in 96-well plates because it was extremely easy to dislodge the cells from the bottom of the well when changing solutions. One possible solution is to coat the bottom of the wells with Collagen I.
Are you plating the cells in serum-free media? If so, possibly try plating the cells in serum-containing media, leaving them for 24 hr, then change to serum-free media for 24 hr then begin your experiment. Plating the cells in serum keeps them in a more synthetic phenotype thereby facilitating increased ECM production and deposition.
Hi, thanks for the reply. Yup, i cultured the cell with serum free media. Ok, i will try with all the suggested methods. Thank you so much.
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