Jan 21, 2026

Public workspaceDissociated Cardioids Assembly Using 30% Matrigel Droplets

DOI
https://dx.doi.org/10.17504/protocols.io.3byl489e2vo5/v1
  • Harry Akligoh1
  • 1Northeastern University - Konry lab
Harry Akligoh
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DOI: https://dx.doi.org/10.17504/protocols.io.3byl489e2vo5/v1
Protocol Citation: Harry Akligoh 2026. Dissociated Cardioids Assembly Using 30% Matrigel Droplets. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl489e2vo5/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: In development
We are still developing and optimizing this protocol
Created: January 21, 2026
Last Modified: January 21, 2026
Protocol Integer ID: 239041
Keywords: dissociated cardioids assembly, cardiac organoid, microfluidic way
Abstract
Matured cardiomyocytes are needed for disease modelling and drug screening. In this protocol we describe a microfluidic way of reassembling dissociated cardiac organoids back into cardiac organoids.
Troubleshooting
30% Matrigel Droplet Generation and Dissociated Cardioid Assembly
Thaw frozen stock of dissociated cardioids in a waterbath set to 37C for two minutes. Don't allow it to thaw completely.
Transfer into 15ml Falcon tube and resuspend in 5ml RMPI 1640+B27 supplement.
Centrifuge at 200xg for 5 min and decant the supernatant.
Add 1ml RPMI 1640+B27 to the cell pellet and resuspend.
Measure cardiomyocytes viability by taking 10ul of cell suspension and adding it to 10ul of Trypan blue (1:1). Count using an automated countess 3 haemocytometer.
Prepare 30% Matrigel by diluting down stock Matrigel with RPMI 1640+B27.
Resuspend 2-5 million dissociated cardioids in 30% Matrigel and use for droplet generation.
Generate about 300um droplets off-chip into 1.5ml eppendorf tube prefilled with 100-500ul of RPMI 1640+B27.
Aliquot 2ml of RMPI 1640+B27 into each well of 6-well plate.
Pipette 100ul of 30% Matrigel-cardiomyocytes droplets into each well of the 6-well plate.
Swell the plate gently making sure not to merge the droplets.
Incubate in a 37C/5% CO2 incubator and change media everyday.
Take images and videos everyday beginning Day 0 until Day 10. On each day check for viability by staining with Calcein-AM (Add 1.5ul of Calcein-AM to 1.5ml of RPMI 1640+B27 and use that to stain the droplets for 2 hours)

Schema of encapsulated dissociated cardioids in 30% Matrigel