Exosome Uptake by Adipocytes

Exosomes derived from breast cancer cells were labelled by the cell membrane labelling agent PKH26 (Sigma-Aldrich). After being seeded in 96-well plates and allowed to differentiate, mature 3T3-L1 cells were incubated with labelled exosomes (20 μl/well) for the indicated time. Images were acquired using the Olympus FluoView FV1000.

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Wu Q., Li J., Li Z., Sun S., Zhu S., Wang L., Wu J., Yuan J., Zhang Y., Sun S, & Wang C. (2019). Exosomes from the tumour-adipocyte interplay stimulate beige/brown differentiation and reprogram metabolism in stromal adipocytes to promote tumour progression. Journal of Experimental & Clinical Cancer Research : CR, 38, 223.

Publication 2019

PKH26 FluoView FV1000

3t3 l1 cells Breast cancer Cell membrane Cells Exosomes Pkh26

Corresponding Organization : Wuhan University

Other organizations : Renmin Hospital of Wuhan University

Top 5 similar protocols

Variable analysis

independent variables

Incubation time of mature 3T3-L1 cells with labelled exosomes

dependent variables

Uptake of labelled exosomes by mature 3T3-L1 cells

control variables

Mature 3T3-L1 cells
Breast cancer cell-derived exosomes
PKH26 labelling agent
96-well plate format
Olympus FluoView FV1000 imaging system

positive controls

Not specified

negative controls

Not specified

Annotations

Based on most similar protocols

Labeling exosomes with PKH26 dye is a common method used across the protocols to track the uptake of exosomes by target cells (Input protocol, Protocol 3).

Exosomes can be isolated from cell culture medium or serum samples using precipitation kits like ExoQuick (Protocol 1). This method provides a relatively simple and fast way to purify exosomes.

Washing the labeled exosomes with PBS after labeling is important to remove any residual dye that could interfere with the experiments (Protocol 3).

Centrifugation at high speeds (e.g., 100,000 × g for 1 hour) can be used to further purify the labeled exosomes and remove any remaining dye (Protocol 3).

The concentration of exosomes used for treating cells is typically in the range of 20 μg/mL to 50 μg/mL (Protocols 2 and 4).

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

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