Dear researchers kindly let me know is it will be feasible to culture blood cells in animal cell culture lab and put the isolated cells to grow in cell culture medium in the same CO2 incubator in which cancer cells are growing? Or it will create problems or have to put separately?
Needs your valuable suggestions
Hi Upadhyay,
Greetings to you. Did you planning to isolate the human monocytes from healthy blood donors and culture it? Do you want to use it as isolated monocytes or as macrophages?
I think for the primary cells such as isolated monocytes should be use separate cell culture hood and for the cell line us other one. Most lab research recommend this style and according to my experience. For the isolated monocytes you can use " RPMI 1640 Medium (Gibco, ThermoFisher, Paisley, UK) supplemented with 1% L-glutamine (Gibco), 1% penicillin/streptomycin (Gibco) and 5% de-complemented human serum" at 37 °C with 5% CO2.
For more information you can read the details of my paper for monocytes and HTP-1.
IJMS | Free Full-Text | Monocytes Expose Factor XIII-A and Stabilize Thrombi against Fibrinolytic Degradation (mdpi.com)
Please don't hesitate to contact me for any further information or help.
Good luck,
Fahad
Blood cells and cancer cells have different growth requirements and sensitivities to environmental conditions. Co-culturing them in the same CO2 incubator can lead to unintended interactions, compromising the physiological relevance of experimental results. Additionally, it increases the risk of cross-contamination, which can impact the reproducibility and reliability of research findings.
Dear Dr. Upadhyay,
You may culture blood cells in animal cell culture lab and use the same CO2 incubator which contains other mammalian cultures namely, cancer cell lines.
I have cultured PBMCs using the same biosafety hood which was used for other mammalian cell lines including cancer cell lines. But you need to avoid cross contamination. So, work with one culture at a time in the laminar hood, and if you have enough space in the incubator, you may place the cultures distant apart, if possible, use a separate rack in the same CO2 incubator for the monocyte culture.
Regards,
Malcolm Nobre.
As mentioned by others, all cells in the same incubator must be able to grow at the same percentage of CO2. This is mainly a function of the culture media and media can often be selected or adjusted as needed.
If you are concerned about cross-contamination by mycoplasma, yeast, bacteria, or viruses between flasks of cultured cells, this is a valid concern. Primary cultures, i.e. cultures started from the cells or tissues obtained directly from living organisms, are much more liable to bring microbial contaminants into cell culture than are continuously cultured cell lines. However, contaminants from any infected primary or continuously cultured cells can be transmitted to other flasks or plates in the same incubator. Using flasks with filter caps can minimize these risks but not entirely prevent cross-contamination. Note that filter capped flasks cannot prevent viruses from passing into or out of the flasks. Because of this, it is always best to use screened, virus-free cells or cultures, if possible and to minimize the production of foam or bubbles in culture media.
In my lab, we minimize cross-contamination by:
1. Never growing continuous cell lines in the same incubator with newly established primary cells.
2. Always using filter capped flasks to reduce cross contamination.
3. Avoiding the use of unfiltered Petri-type or multi-well plates in the same incubator as continuous cell lines.
Best of luck,
RW
I would add a couple of comments to the response by Robert J Walter. In my experience the primary source of cross contamination is feeding, trypsinizing, washing etc different cultures from the same media/trypsin/HBSS bottles. You should always have a different bottle of media etc, even if the same type of media for each cell line/primary culture. In addition, always change pipettes etc between different cell cultures. Of course if you get mycoplasma contamination, kill everything in the incubator, sterilize the incubator, throw out all media, trypsin and saline etc and start over.
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