Exosome spin columns mw 3000

Manufactured by Thermo Fisher Scientific

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About the product

The Exosome Spin Columns (MW 3000) are a laboratory tool designed for the isolation and purification of exosomes from various biological samples. These columns utilize a molecular weight cut-off of 3000 Da to selectively retain exosomes while allowing smaller molecules to pass through. The columns are suitable for use with a range of sample types, including cell culture supernatants, body fluids, and other biological matrices.

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Exosome Spin Columns - 108 citations Spin columns - 107 citations MW 3000 - 42 citations Exosomes - 6 citations

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43 protocols using «exosome spin columns mw 3000»

Tracking Exosome Uptake by BMSCs

2025

To identify whether SCs-derived exosomes could be taken up by BMSCs, the fluorescent dye PKH67 (green) (Sigma-Aldrich, St. Louis, MO) was used to label exosomes according to the manufacturer’s protocol. Briefly, exosomes were extracted from SCs and diluted in PBS to a final concentration of 100 mg/ml, and 2 ml of PKH67 dye was mixed with 250 ml of Diluent C (PHK67 solution) [32 (link), 33 (link)]. Then, 600 ml of the diluted exosomes and 200 ml of the PKH67 mixture were added to a 1.5-ml centrifugation tube and incubated at room temperature for 5 min. Centrifugation at 750 × g for 2 min was performed to remove the unincorporated dye by using exosome spin columns (MW3000; Invitrogen, Vilnius, Lithuania). The labeled exosomes, resuspended in exosome-free DMEM, were incubated with BMSCs for 4 h at 37 °C. Nuclei were stained with DAPI (blue) (Solar Bio, Beijing, China), and F-actin was stained with tetramethylrhodamine isothiocyanate phalloidin (red) (MKBio, Shanghai, China). Images were then captured using a confocal microscope.

Wang Y., Qu F., Wu Y., Lan K., Shen Y., Wu Z., Zhong Q., Cao X., Fan Z, & Xu C. (2025). Peripheral nerves modulate the peri-implant osteogenesis under type 2 diabetes through exosomes derived from schwann cells via miR-15b-5p/Txnip signaling axis. Journal of Nanobiotechnology, 23, 51.

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Modulating CD8+ T Cells with Extracellular Vesicles

2025

For in vitro EVs treatment, a total of 2.5 × 10⁸ EVs (equivalent to those collected from 6 × 10⁶ producer cells, 50 mL culture medium) derived from Hepa1-6-Ctrl or Hepa1-6-HDAC2-AS2 cells were added to 2 × 10⁵ activated CD8⁺ T cells for 48 h, PBS as a control treatment. Then, the cells were collected for western blot analysis, RNA extraction, and flow cytometry detection. For in vitro EVs labeling, DiI (Beyotime Biotechnology, China) was added to PBS buffer or EVs suspension at a concentration of 1 μM and incubated for 30 min at 37 °C, followed by washing through exosome spin columns (MW3000) (Invitrogen) to remove excess dye. DiI-labeled EVs were incubated with activated CD8⁺ T cells for 24 h. As a control, PBS buffer containing free DiI dye was washed through exosome spin columns. In addition, a total of 2 × 10⁸ EVs (equivalent to those collected from 5 × 10⁶ producer cells in 50 mL of culture medium) derived from Hepa1-6-Ctrl-EGFP-mCD63 or Hepa1-6-HDAC2-AS2-EGFP-mCD63 cells were added to 2 × 10⁵ activated CD8⁺ T cells and incubated for 24 h.

Gao Y., Zhang Z., Huang X., You M., Du C., Li N., Hao Y., Wang K., Ding X., Yang F., Cheng S.Q., Luo J., Chen R, & Yang P. (2025). HBV-associated hepatocellular carcinomas inhibit antitumor CD8+ T cell via the long noncoding RNA HDAC2-AS2. Nature Communications, 16, 2055.

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Evaluating Exosome and Mimetic Uptake

2024

To evaluate exosome- and mimetic-mediated cell uptake by human fetal lung fibroblasts (MRC-5) and mouse macrophages (J774), 1×10⁴/cm² MRC-5 and J774 cells were seeded into 6-well plates and allowed to adhere overnight. The next day, exosomes and mimetics were stained with 5 µM Vybrant DiD dye (ThermoFisher) in a final volume of 500 µl for 10 min at 37°C. Washes were performed by ultracentrifuging mixture at 40,000xg for 1 hr at 4°C. The pellet was resuspended in 1 ml of 0.22 µm filtered PBS (Gibco) and exosome spin columns (MW3000, Invitrogen) were used to remove any unbound dye. DiD stained exosomes and mimetics (at the concentration of 1×10⁸) were added to each well. Cellular uptake was quantitatively evaluated at different time points (4, 8, 12 hrs) by fluorescence microscopy. The efficiency of exosomes and mimetics in protecting mRNA while keeping it functional, MRC-5 and J774 were exposed to mRNA-loaded particles for 24, 48 and 72 hrs. At the end of each incubation cells were processed for flow cytometry. The same approach was followed to test the efficacy of F-MIMs as delivery systems.

Fonteles C.S., Steele J.W., Idowu D.I., Burgelin B., Finnell R.H, & Corradetti B. (2024). Amniotic fluid-derived stem cells: potential factories of natural and mimetic strategies for congenital malformations. Research Square.

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Internalization of Exosomes by hBMSCs

2024

HBMSCs were purchased from Cyagen Biosciences. To detect whether Exos could be internalized by HBMSCs, PM-Exos, and OM-Exos were labeled by Dil (Beyotime Biotechnology, China), a lipophilic fluorescent membrane dye for easy tracking. Initially, Dil was diluted to a final concentration of 10 μM and incubated with Exos for 30 min at 37 °C. Exosome Spin Columns MW3000 (Thermo Fisher Scientific, USA) were used to remove the unincorporated dye. After that, labeled Exos of 10¹⁰ particles/ml were co-cultured with HBMSCs for 4 h. Subsequently, the supernatant was discarded and HBMSCs were washed twice with 10% PBS to remove free Exos and fixed in 4% PFA for 10 min. Hoechst 33342 (Beyotime Biotechnology, China) and Alexa Fluor 488 phalloidin (Thermo Fisher Scientific, USA) were separately used for the staining of nuclei and F-actin. The images were obtained under a confocal laser scanning microscope (CLSM, Leica, Germany). In the blank control group, Exos were replaced by PBS to co-culture with HBMSCs.

Li S., Rong Q., Zhou Y., Che Y., Ye Z., Liu J., Wang J, & Zhou M. (2024). Osteogenically committed hUCMSCs-derived exosomes promote the recovery of critical-sized bone defects with enhanced osteogenic properties. APL Bioengineering, 8(1), 016107.

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Exosome Uptake by Cancer Cells

2023

Exosomes were collected by density gradient (1.15–1.19 g/mL) ultracentrifugation according to previously published protocol.^{15 (link)} After ultracentrifugation preparations, exosomes were labeled with PKH67 Fluorescent Cell Linker Kits (Sigma-Aldrich) according to the manufacturer’s instructions, followed by washing through Exosome Spin Columns (MW3000) (Invitrogen) to remove excess dye. Next, 5 μg exosomes were incubated with 1 × 10^{5 (link)} MFC or other cancer cells for 24 h which were examined under a confocal microscope at the indicated time points.

Zheng N., Wang T., Luo Q., Liu Y., Yang J., Zhou Y., Xie G., Ma Y., Yuan X, & Shen L. (2023). M2 macrophage-derived exosomes suppress tumor intrinsic immunogenicity to confer immunotherapy resistance. Oncoimmunology, 12(1), 2210959.

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