I have a question on cultured cell staying in RPMI medium for a month without an idea what is it.
Some informations are;
Source of sample: blood sample from cancer patient.
Separation method: electrical field.
Cell culture: Isolated cells have been cultured with RPMI1640 + 20%FBS + antibiotics.
My guess is that blood cells should not attach to Well Plate and can be skimmed off. The attached cell could be epithelial or endothelial cells?
Do you have access to a microscope with a scale? If the cell is big (>10-20 um) and has been sitting there for a month doing nothing, I would guess macrophage. Second guess, endothelial cell.
Its for sure not epithelial as most will not grow in RMPI and they grow in form of colonies. Looking at those little projections comming out from the sides I feel its a macrophage.
For me it is looking like mesenchymal cells, because of large size, projections help them in migration. The nucleus size is small but in macrophage it should be little bit larger. In macrophage the vacuoles should be present in higher frequency than what is appearing here. In endothelial cells less projections are there than what we can see here, it only appears when special media with growth factors provided. Taken together I think it is EMT (epithelial to mesenchymal transition) product.
I would go for macrophage as well, definitely not a mesenchymal cells.
Many years ago I studied the tooth granulomas and find in most of them macrophages in end stage of their life, accumulating fat and appearing as foam cells. Foam cells are characteristic for atheromatous plaques and are macrophages with the same mophology, loaded with cholesterol.Your cell is an old macrophage undergoing fat degeneration.Macrophages are mesodermal derivatives originating in the serous cavities.When they enter by diapedesis the blood circulation, they round up and became monocytes. They are attracted to areas in inflammation, destinguishing self from nonself. They help by cleaning the field by phagocytosis, transmitt the informaton to lymphocytes and finally turn into foam cells.
Please see " Origin and Development of the Immune system" in my publications.
May be a monocyte/macrophage, but you need to perform ICC with specific markers as CD11b, CD14.The cultured mesenchymal cells express other surface markers, while lacking the expression of CD11b, CD14.
Hi, you do not mention if these cells are dividing or not. Macrophages do not normally divide in culture, unless stimulated by specific growth factors. I agree with Gabriel Bonaterra, and suggest performing immunophenotyping (if the cells divide).
Sittisak, I'm curious: what about your cells? Did you recognise them? Are they still alive?
Dear Sittisak,
I think endothelial cell. Since you can see some ciliated structure at the anterior and basolateral side.
Best,
Monalisa
The fact that the cell is not dividing and die approve my opinion, that it is an old macrophage undergoing fat degeneration.
Hello all friends here,
Currently I have been culturing isolated CTCs from blood of cancer patient. Isolated cells are growing on Matrigel coated on bottom of 24-well plate. My idea is to have a kind of ECM (Extracellular Matrix) helping cells to healthy grow. So far (almost 1 month after isolation) I can observe serveral cells attach on Matrigel which is better result than without Matrigel coated. Please see images of them with this message and share any comments what they are?
Hi,
Did your CTCs divide on matrigel?
Could you share with us some CTCs images before culturing on matrigel?
I'm sorry but to me these look like dying cells Some are detached and dead and attached cell internal structures looks like apoptosis in progress because there is no obvious nucleus.
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