Non-destructive in situ monitoring of structural changes of 3D tumor spheroids during the formation, migration, and fusion process
- Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, China
- Key Laboratory of Animal Biological Products & Genetic Engineering, Ministry of Agriculture and Rural, Sinopharm Animal Health Corporation Ltd, China
- State Key Laboratory of Novel Vaccines for Emerging Infectious Diseases, China National Biotec Group Company Limited, China
- School of Computer and Information Science, Southwest University, China
Figures
Figure 1
Validation of the non-destructive in situ observation of three-dimensional (3D) spheroids by the side-view observation device.
(A) The illustration of the side-view observation device. (B) Exploded view diagram of the side-view observation device. (C) The definition of the bottom- and the side-view of a spheroid sample. (D) …
Figure 2
Spheroids’ observation and imaging setup.
(A) Light path of the bottom-view observation of the spheroid in the agarose micro-well. (B) Light path of side-view observing of the spheroid in the agarose micro-well. (C) The absorption spectrum …
Figure 3
The formation dynamics of the spheroids.
(A) Time-lapse images of DU 145 spheroid formation in an agarose well (1 × 104 cells/well, scale bar = 100 μm). (B) Changes in height and width during the growth process of the spheroids, as well as …
Figure 4
The migration of the spheroids.
(A) The bottom-view images of the spheroid in the petri dish, cells extrude from the spheroid, forming sprouts. (B) Statistics on the changes in total width, inner width, and migration distance of …
Figure 5
The dynamics of dark zone formation within the three-dimensional (3D) spheroid.
(A) Time-lapse bottom- and side-view images of DU 145 spheroids formed from different initial cell concentrations (2.5 × 103, 5 × 103, 1 × 104, and 2 × 104 cells/well). Blue arrows indicate the …
Figure 6
Characterization of the spheroids fusion process.
(A) Time-lapse bottom- and side-view images of fusion body formed by two 7-day-old DU 145 cell spheroids with ideal ball-like structure (i) and irregular cell spheroid (ii). Red arrows point to the …
Figure 7
The in situ observation of DU 145 spheroids under the killing effect of doxorubicin (DOX) and natural killer (NK) cells.
(A) Time-lapse bottom- and side-view images of 3- and 7-day-age DU 145 cell spheroids (1 × 104 cells/spheroid) under DOX (50 μg/mL) treatment. (B) The changes of DU 145 spheroids’ width and height …
Figure 8
The display of the side-view observation petri dish/device.
(A) The top- and side-view images of the side-view observation petri dish/device. (B) The magnets inside the device.
Author response image 1
The device was used on an inverted microscope to capture fluorescence images.
(A) Schematic of the imaging setup. (B) Bright-field and fluorescence images of cells. (Scale bar = 500 µm.)
Videos
Video 1
Observation of the bottom- and side-view of a tumor spheroid using the side-view observation device.
Video 2
Locating the side-view of the spheroid in the agarose micro-well through the bottom-view.
Video 3
Recording the side-view images of different spheroids in the agarose micro-well.
Video 4
Non-destructive in situ observation of the thickness changes during DU 145 cell spheroids formation process.
Video 5
Non-destructive in situ observation of the thickness changes during DU 145 cell spheroids migration process.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (human prostate cancer cell) | DU 145 | Pricella Biotechnology | CL-0075 (RRID:CVCL_0105) | |
| Cell line (human natural killer cell) | NK-92 | Pricella Biotechnology | CL-0530 (RRID:CVCL_2142) | |
| Chemical compound, drug | DMEM culture medium | Gibco | C11995500BT | |
| Chemical compound, drug | Fetal bovine serum | Bio-channel | BC-SE-FBS01 | |
| Chemical compound, drug | Penicillin–streptomycin solution | Beyotime Biotechnology | C0222 | |
| Chemical compound, drug | NK-92 special medium | Pricella Biotechnology | CM-0530 | |
| Chemical compound, drug | Agarose | Aladdin Scientific | 11966311 | |
| Chemical compound, drug | Phosphate-buffered saline | Beijing DingGuo ChangSheng Biotechnology Co Ltd | BF-0011 | |
| Chemical compound, drug | Ultrapure water | ELGA Corporation | PURELAB flex system | |
| Software, algorithm | Origin | OriginLab Corporation | 2021 (RRID:SCR_014212) | |
| Software, algorithm | ImageJ | LOCI, University of Wisconsin | (RRID:RCR_003070) | |
| Other | Xenon lamp | Beijing zhongjiao Jinyuan Technology Co, Ltd | CEL-TCX250 | Light source |
| Other | Fiber optic spectrometer | Fuxiang Optics Co, Ltd | PG2000-Pro-EX | Instrument |
| Other | Microscope | Nikon | ECLIPSE Ti (RRID:SCR_021242) | Instrument |
| Other | D-LH/LC lamphouse | Nikon | Light source | |
| Other | 3D printer | Bambu Lab | P1P | Instrument |
