I'm going to transfect cells with expression plasmids and later siRNAs against the same gene. Is it true that penicillin/streptomycin in the media negatively affects these processes? Shout I use media without antibiotic even when I'm seeding the cells?
Thank you for your help in advance!
Hello Bahar Ksh
Generally, antibiotics can be present in the medium for transient transfection. However, when cationic lipid reagents are used, cell permeability increases and this may also increase the amount of antibiotics that is delivered into the cells, resulting in cytotoxicity and lower transfection efficiency. So, it is better not to use antibiotics in the transfection medium unless previously tested in the cell type being transfected.
Also, avoid antibiotics when plating the cells for transfection as this will reduce the need for rinsing the cells before transfection.
In cell culture avoid the use of antibiotics as far as possible. If you maintain good sterile conditions, you may discontinue the use of antibiotics in culture.
Best.
I am not an expert in this field, but I am very interested and have researched to find an answer. Could you please review the response below to see if it is correct?
The use of antibiotics such as penicillin and streptomycin in cell culture media can negatively impact transfection processes. Research indicates that these antibiotics interfere with cell differentiation and growth rates. Specifically, studies have shown that penicillin-streptomycin can reduce the expression of specific markers and the growth rate of differentiating cells by significant percentages [1][2]. Additionally, the use of antibiotics is unnecessary if aseptic techniques are properly followed [1]. Therefore, it is advisable to avoid using media containing antibiotics during transfection procedures to maximize efficiency and accuracy. This recommendation applies even when seeding the cells, to prevent any adverse effects from the outset.
Reference
[1] Llobet, L., Montoya, J., López‐Gallardo, E., & Ruiz‐Pesini, E. (2015). Side Effects of Culture Media Antibiotics on Cell Differentiation.. Tissue engineering. Part C, Methods, 21 11, 1143-7 .
[2] Cohen, S., Samadikuchaksaraei, A., Polak, J., & Bishop, A. (2006). Antibiotics reduce the growth rate and differentiation of embryonic stem cell cultures.. Tissue engineering, 12 7, 2025-30 .
I received some assistance from tlooto.com for this response.
From my personal experience, it depends on the cell line and transfection reagent. As a general rule, avoid antibiotics for transfection but I routinely transfect HEK293T cells with either PEI or calcium phosphate in medium + ab with no obvious decrease in transfection efficiency.
Hi Park Sowon ,
I'm also no experts here, but I've been using pen/strp for many different adherent and non-adherent cells for a few years and never had any issues with decreased growth. Also while the pathway that I'm working on is immune-related, I haven't had any issues with altered gene expression as well.
But I think all this depends on the cell line you are working with and how sensitive/tolerant they are.