Feb 13, 2026
In vitro LDTIA of tumors based on MTS formation
- Guangli Suo1
- 1CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Jiangsu, 215123, China.
- MTS
Protocol Citation: Guangli Suo 2026. In vitro LDTIA of tumors based on MTS formation. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk12kwg5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 13, 2026
Last Modified: February 13, 2026
Protocol Integer ID: 243173
Keywords: ldtia of tumor, clinical brc tissue, brc tissue, proportion of csc, tumor, csc, vitro model
Funders Acknowledgements:
National Natural Science Foundation of China
Grant ID: 32470814
Abstract
A in vitro model for determining the proportion of CSCs in clinical BRC tissue.
Troubleshooting
Four microengineered MWC-chips with microwells of varying diameters (80µm, 50µm, 30µm, and 15µm) and uniform depth (40µm) were designed to accommodate approximately 100, 40, 10, and 1 tumor cells per microwell, respectively. Tumor tissues from clinical BRC patients were dissociated into single-cell suspensions by enzymatic digestion and seeded onto the four different-sized MWC-chips for MTS generation.
MFR was calculated from 108 microwells in one microscopic field as: MFR = (number of MTSs / total microwells) × 100%.
The CSC frequencies were determined following the previous reported method. Briefly, a table listing the number of microwells containing MTSs and the corresponding cell count per microwell for each chip type was prepared.
Data were input into the online tool at http://bioinf.wehi.edu.au/software/elda/, which outputs a table with the estimated tumor-initiating cell frequency and its 95% confidence interval, along with a log-fraction plot of the limiting dilution model showing cell number per well (x-axis) versus nonresponders.