Overview

Norgen's FBS Exosome Depletion Kits provides a quick and easy protocol for the depletion of bovine exosomes from FBS prior to using it as a growth supplement in your culture medium. The FBS recovered from the depletion process is exosome-depleted and does not contain any quantifiable bovine miRNAs. Moreover, the exosome-depleted FBS will support the growth of your cells of interest similar to the non-depleted FBS. Norgen's kits allow for the processing of different FBS volumes. The depletion is based on Norgen’s proprietary resin. These kits provide a clear advantage over other available kits in that they do not require ultracentrifugation, any special instrumentation, precipitation reagents or any protease treatments. More importantly, the depletion process is an inexpensive method for exosome depletion from FBS, as compared to the expensive current ready-to-use exosome-depleted media available on the market.

Background

Most culture medium used for the growth and propagation of cells in culture require the addition of fetal bovine serum (FBS) as a growth supplement to media. FBS is obtained from bovine (cow) serum, and therefore contains large quantities of bovine exosome vesicles. These exosomes may interfere with some types of studies, or may lead to unreliable results when studying the exosomes shed from your cells of interest in normal culture conditions. Therefore, the use of exosome-depleted FBS is highly recommended for many types of studies.

FBS Exosome Depletion Kit (Slurry Format)

For FBS volumes ranging from 140 mL to 280 mL.

FBS Exosome Depletion Kit (Column Format)

For FBS volumes ranging from 120 mL to 240 mL.

Details

Supporting Data

Previous

Figure 1. Exosome-depleted FBS with Norgen's FBS Exosome Depletion Kits (Slurry Format) has undetectable Bovine miRNA levels. Norgen's FBS Exosome Depletion Kit I (Slurry Format) (Cat# 61100) was used to deplete bovine exosomes from 5mL FBS. Total RNA/miRNA including exosomal RNA was purified from the depleted FBS, non-depleted FBS and a commercially available ready – to – go depleted FBS using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit (Cat# 55800). Five different bovine microRNAs were assessed by RT-qPCR (miR-26a, miR-30a, miR-92a, miR-23a and miR-122). Three out of the five tested miRNA (miR-26a, miR-92a and miR-23a) didn’t show any amplification in the FBS depleted using Norgen's FBS Exosome Depletion Kit I (Slurry Format) whereas the other two miRNAs (miR-30a and miR-122) showed very late Ct. values which appeared to be a primer dimer according to the melt curve.

Figure 2. Growth rates of HeLa cells in media containing Exosome-depleted FBS. Growth rates of HeLa cells in media containing Exosome-depleted FBS using Norgen's FBS Exosome Depletion Kits (Slurry Format) was compared to that in media containing standard FBS. Simply, HeLa cells were seeded in DMEM with either 10% Exosome-depleted FBS using Norgen's Kits or 10% standard FBS and then cultured under standard conditions at 37°C with 5% CO² for 3 days. The cells were imaged using Moticam 480 to observe cellular morphology and growth rate. Similar growth and identical cellular morphology were detected for both the Exosome-depleted FBS using Norgen’s FBS Exosome Depletion Kits and the standard FBS

Figure 3. Exosome-depleted FBS with Norgen's FBS Exosome Depletion Kits (Column Format) has undetectable Bovine exosomes levels. Norgen's FBS Exosome Depletion Kit I (Column Format) (Cat# 61200) was used to deplete bovine miRNA from 5mL FBS. Total RNA/miRNA including exosomal RNA was purified from the depleted FBS, non-depleted FBS and a commercially available ready – to – go depleted FBS using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit (Cat# 55800). Five different bovine microRNAs were assessed by RT-qPCR (miR-26a, miR-30a, miR-92a, miR-23a and miR-122). Three out of the five tested miRNA (miR-26a, miR-92a and miR-23a) didn’t show any amplification in the FBS depleted using Norgen's FBS Exosome Depletion Kit I (Column Format) whereas the other two miRNAs (miR-30a and miR-122) showed very late Ct. values which appeared to be a primer dimer according to the melt curve.

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