In vitro nanoparticle uptake assays

David T. Omstead; Franklin Mejia; Jenna Sjoerdsma; Baksun Kim; Jaeho Shin; Sabrina Khan; Junmin Wu; Tanyel Kiziltepe; Laurie E. Littlepage; Basar Bilgicer

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In vitro nanoparticle uptake assays

DO David T. Omstead

FM Franklin Mejia

JS Jenna Sjoerdsma

BK Baksun Kim

JS Jaeho Shin

SK Sabrina Khan

JW Junmin Wu

TK Tanyel Kiziltepe

LL Laurie E. Littlepage

BB Basar Bilgicer

This method is extracted from research article: J Hematol Oncol, Nov 2020

In vivo evaluation of CD38 and CD138 as targets for nanoparticle-based drug delivery in multiple myeloma

DOI: 10.1186/s13045-020-00965-4

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Per well, 100,000 cells were plated in a 24-well dish 6 h prior to each experiment. Nanoparticles were prepared with DiD as a fluorescent maker as described in the section that explains nanoparticle preparation and added to the wells at a 100 µM phospholipid concentration and incubated at 37 °C for 1 to 24 h as described in each experiment. After incubation, cells were washed twice with PBS and then incubated for 10 min with 100 µL of 0.25% Trypsin–EDTA solution (Sigma-Aldrich) to cleave off nanoparticles bound to the surface of the cell but not yet endocytosed. After trypsinization, cells were washed once more with PBS and then analyzed via flow cytometry.

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