Spheroid surgery: a first-generation micromanipulation platform for extracting viable cell clusters from the outer layers of hydrogel-embedded spheroids
- 1HUN-REN Biological Research Centre
Abstract
Three-dimensional (3D) cell culture systems have gained attention as in vitro models capable of reproducing the tissue-like architecture, spatial organization, and physiological morphology of in vivo tumors. Their ability to mimic native cell-cell and cell-matrix interactions has positioned 3D cell cultures as powerful models for studying tumor biology and drug responses. However, methods for the localized extraction of viable cells remain limited, as existing approaches often disrupt the original 3D structure, limit access to defined regions, or compromise cell viability, restricting the biological insight that can be achieved. We present a manual, micropipette-based manipulation system designed to extract viable multicellular clusters from the outer layers of compact 3D tumor spheroids, embedded in biocompatible hydrogels. The extracted clusters remained fully functional, demonstrating that the procedure is sufficiently gentle to remove small multicellular units. This technology enables the localized extraction of viable multicellular clusters from 3D in vitro models without complete dissociation of the parent spheroid. The method opens new opportunities for longitudinal studies, spatially targeted sampling and controlled manipulation of living 3D cultures. This capability marks a step toward a new era of 3D model manipulation, where researchers can add, remove, or replace specific regions of living multicellular systems with precision.
Related
- Microfluidic generation of alginate microbeads containing in situ-formed apical papilla stem cell spheroids and NecroX-5-loaded nanoparticles
- A modular synthetic DNA template platform for rapid generation of TCR-encoding mRNA and streamlined validation of cognate TCR–epitope reactivity
- High efficiency gene editing and phenotype generation in avian embryos using lipid nanoparticles as a vector delivery platform
- A pathway to next-generation mast cell stabilizers identified through the novel Phytomedical Analytics for Research Optimization at scale data platform.
- A quantitative three-dimensional bone mineralisation assay using human dental pulp stem cell spheroids
BibTeX
@misc{pal2026spheroid,
title = {{Spheroid surgery: a first-generation micromanipulation platform for extracting viable cell clusters from the outer layers of hydrogel-embedded spheroids}},
author = {V. Pal and M. Maiurov and M. Stellato and N. Hapek and M. Harmati and F. Piccinini and A. Diosdi and P. Horvath},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.22.760885},
url = {https://www.biorxiv.org/content/10.64898/2026.10.22.760885v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Spheroid surgery: a first-generation micromanipulation platform for extracting viable cell clusters from the outer layers of hydrogel-embedded spheroids
AU - V. Pal
AU - M. Maiurov
AU - M. Stellato
AU - N. Hapek
AU - M. Harmati
AU - F. Piccinini
AU - A. Diosdi
AU - P. Horvath
PY - 2026
DA - 2026/10/22
PB - bioRxiv
DO - 10.64898/2026.10.22.760885
UR - https://www.biorxiv.org/content/10.64898/2026.10.22.760885v1
AB - Three-dimensional (3D) cell culture systems have gained attention as in vitro models capable of reproducing the tissue-like architecture, spatial organization, and physiological morphology of in vivo tumors. Their ability to mimic native cell-cell and cell-matrix interactions has positioned 3D cell cultures as powerful models for studying tumor biology and drug responses. However, methods for the localized extraction of viable cells remain limited, as existing approaches often disrupt the original 3D structure, limit access to defined regions, or compromise cell viability, restricting the biological insight that can be achieved. We present a manual, micropipette-based manipulation system designed to extract viable multicellular clusters from the outer layers of compact 3D tumor spheroids, embedded in biocompatible hydrogels. The extracted clusters remained fully functional, demonstrating that the procedure is sufficiently gentle to remove small multicellular units. This technology enables the localized extraction of viable multicellular clusters from 3D in vitro models without complete dissociation of the parent spheroid. The method opens new opportunities for longitudinal studies, spatially targeted sampling and controlled manipulation of living 3D cultures. This capability marks a step toward a new era of 3D model manipulation, where researchers can add, remove, or replace specific regions of living multicellular systems with precision.
ER -