Table 1 Specifications of experimental setups according to MISEV 2018,2023

Pre-processing variables

 Isolation source

Conditioned medium from cell culture

 Cells

AdSCs

 Provenance

Primary cells

 Time of culture and confluency

3–4 days, 70–80% confluency

Culture and EV isolation conditions

 Passage

2–3

 Culture volume

20ml

 Culture vessel

T175cm² flasks

 Surface coatings

none

 Culture conditions

5% CO_2, 37°C, 95% H₂O

 Conditioning medium

DMEM (4.5mg/l D-glucose), 1% P/S, 2ng/ml b-FGF

 Collection method

Pipetting from flasks

Isolation

 Differential centrifugation with filtration step

300xg 10 min 4°C

2,000xg 20 min 4°C

200nm PVDF Filter

2 × 100,000xg 1,5h, 4°C

Resuspension volume 100µl of PBS

Storage conditions

− 80°C in PBS and low protein binding Eppendorf tubes