Non-destructive in situ monitoring of structural changes of 3D tumor spheroids during the formation, migration, and fusion process

  1. Ke Ning
  2. Yuanyuan Xie
  3. Wen Sun
  4. Lingke Feng
  5. Can Fang
  6. Rong Pan
  7. Yan Li  Is a corresponding author
  8. Ling Yu  Is a corresponding author
  1. 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
  2. Key Laboratory of Animal Biological Products & Genetic Engineering, Ministry of Agriculture and Rural, Sinopharm Animal Health Corporation Ltd, China
  3. State Key Laboratory of Novel Vaccines for Emerging Infectious Diseases, China National Biotec Group Company Limited, China
  4. School of Computer and Information Science, Southwest University, China
9 figures, 5 videos, 1 table and 1 additional file

Figures

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) …

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 …

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 …

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 …

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 …

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 …

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 …

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 resourceDesignationSource or referenceIdentifiersAdditional information
Cell line (human prostate cancer cell)DU 145Pricella BiotechnologyCL-0075 (RRID:CVCL_0105)
Cell line (human natural killer cell)NK-92Pricella BiotechnologyCL-0530 (RRID:CVCL_2142)
Chemical compound, drugDMEM culture mediumGibcoC11995500BT
Chemical compound, drugFetal bovine serumBio-channelBC-SE-FBS01
Chemical compound, drugPenicillin–streptomycin solutionBeyotime BiotechnologyC0222
Chemical compound, drugNK-92 special mediumPricella BiotechnologyCM-0530
Chemical compound, drugAgaroseAladdin Scientific11966311
Chemical compound, drugPhosphate-buffered salineBeijing DingGuo ChangSheng Biotechnology Co LtdBF-0011
Chemical compound, drugUltrapure waterELGA CorporationPURELAB flex system
Software, algorithmOriginOriginLab Corporation2021 (RRID:SCR_014212)
Software, algorithmImageJLOCI, University of Wisconsin(RRID:RCR_003070)
OtherXenon lampBeijing zhongjiao Jinyuan Technology Co, LtdCEL-TCX250Light source
OtherFiber optic spectrometerFuxiang Optics Co, LtdPG2000-Pro-EXInstrument
OtherMicroscopeNikonECLIPSE Ti (RRID:SCR_021242)Instrument
OtherD-LH/LC lamphouseNikonLight source
Other3D printerBambu LabP1PInstrument

Additional files

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