Exosome Uptake by Cancer Cells

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.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

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.

Publication 2023

PKH67 Fluorescent Cell Linker Kits Exosome Spin Columns (MW3000)

Cancer Cells Confocal microscope Exosomes Pkh67 Ultracentrifugation

Corresponding Organization : Shanghai Jiao Tong University

Other organizations : Renji Hospital, First Affiliated Hospital Zhejiang University, Dongguan People’s Hospital, Southern Medical University

Top 5 similar protocols

Variable analysis

independent variables

Density gradient (1.15–1.19 g/mL) ultracentrifugation for exosome collection
Labeling of exosomes with PKH67 Fluorescent Cell Linker Kits
Incubation of 5 μg of labeled exosomes with 1 × 10^5 MFC or other cancer cells for 24 h

dependent variables

Exosome uptake by MFC or other cancer cells examined under a confocal microscope at the indicated time points

control variables

Exosome collection protocol as previously published
Manufacturer's instructions for PKH67 Fluorescent Cell Linker Kits
Exosome Spin Columns (MW3000) used to remove excess dye

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

Based on most similar protocols

The protocols use density gradient ultracentrifugation (1.15–1.19 g/mL) to collect exosomes, which is a common and well-established technique (Input protocol).

The exosomes are labeled with fluorescent dyes (PKH67, PKH26) to allow for visualization and tracking of their uptake by target cells (Input protocol, Protocol 2, Protocol 3).

Exosomes are washed using Exosome Spin Columns or through additional ultracentrifugation steps to remove excess dye (Input protocol, Protocol 2).

The concentration of exosomes is quantified using either the BCA method or by measuring protein content (Protocol 4, Protocol 5).

Nanoparticle tracking analysis (NTA) is used to characterize the size distribution of the isolated exosomes (Protocol 2).

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!