How to remedy cell contamination during cell culture flask culture

Wed Aug 17 11:29:56 CST 2022

When using cell culture flasks to culture cells, sometimes cell contamination occurs. How to remedy this time?

We immediately discarded the medium in the cell culture flask, rinsed 2~3 times with PBS, added trypsin until the cell morphology changed slightly but did not leave the container, discarded the trypsin, and added PBS to gently rinse once;

Add 20× double antibody to the cell culture flask, soak the cells for 3~5min, discard the double antibody, and add fresh complete medium (containing 10× double antibody) for 12h;

cell culture flask

        FuDau T25 Cell Culture Flasks

After 12 hours, discard the medium and wash it with PBS for 2~3 times, then add complete medium (containing 10× double antibody), and centrifuge at a small speed during passage (if centrifugation is usually performed at 1000r/min, it can be reduced to 800~900r/min). min centrifugation), and cultured in medium containing 10× double antibody.

If no bacteria can be found under the second generation rear microscope, the third generation can be cultured normally. If there is no rebound after 48h of normal culture, the contamination can be considered to have been cleared.

In case of mold contamination, change the medium after rinsing with PBS for 2 to 3 times, without adding high-concentration double antibody. After culturing for 48h, the contamination did not reappear.

cell culture flask

     FuDau T75 Cell Culture Flasks

In case of fungal contamination, amphotericin B can be added after rinsing with PBS and changing the medium (Amphotericin B is cytotoxic and not recommended), or 300 μg/mL fluconazole can be added for culture, and 150 μg can be used after contamination control. /mL cultured for 2~3 passages.

If mycoplasma contamination is suspected, PCR detection can be performed or a mycoplasma removal reagent (eg, a Hengbi) can be used directly. Ciprofloxacin injection can also be purchased, opened in the ultra-clean bench and added directly to the culture medium. After 2 generations at a concentration of 20 μg/mL, the concentration of 10 μg/mL was used for continuous cultivation for about 2 weeks.

The above is how to remedy cell contamination when using cell culture flasks to culture cells.

The FAI climbed 5.9 percent year-on-year in the first 11 months of 2018, quickening from the 5.7-percent growth in Jan-Oct, the National Bureau of Statistics (NBS) said Friday in an online statement.

The key indicator of investment, dubbed a major growth driver, hit the bottom in August and has since started to rebound steadily.

In the face of emerging economic challenges home and abroad, China has stepped up efforts to stabilize investment, in particular rolling out measures to motivate private investors and channel funds into infrastructure.

Friday's data showed private investment, accounting for more than 60 percent of the total FAI, expanded by a brisk 8.7 percent.

NBS spokesperson Mao Shengyong said funds into weak economic links registered rapid increases as investment in environmental protection and agriculture jumped 42 percent and 12.5 percent respectively, much faster than the average.

In breakdown, investment in high-tech and equipment manufacturing remained vigorous with 16.1-percent and 11.6-percent increases respectively in the first 11 months. Infrastructure investment gained 3.7 percent, staying flat. Investment in property development rose 9.7 percent, also unchanged.