isoCell User Experience Summary

Single-Cell Cloning Automation Technologies
Overview
Independent scientists from multiple research labs and core facilities have come together toshare their new protocols for improving clone isolation and expansion of human pluripotent stem
cells (hPSCs). Their detailed protocols and user experiences, utilizing emerging single-cell cloning
technologies, are presented in an open access technical paper published in the Wiley Online
Library - Current Protocols in Stem Cell Biology e123, Volume 55, 2020.
Title: Methods for Automated Single Cell Isolation and Sub-Cloning of Human Pluripotent Stem
Cells
Authors: Valeria Fernandez Vallone1,2, Narasimha Swamy Telugu3,4, Iris Fischer1,2, Duncan Miller3,4,5,Sandra Schommer3,4, Sebastian Diecke3,4,5 and Harald Stachelscheid1,2.
Affiliations

Charite - Universitätsmedizin Berlin, Berlin, Germany.
Berlin Institute of Health (BIH), BIH Stem Cell Core Facility, Berlin, Germany.
3
Max Delbruck Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
4
BIH Stem Cell Core Facility, Berlin Institute of Health (BIH), Berlin, Germany.
5
DZHK (German Centre for Cardiovascular Research), partner site Berlin, Berlin, Germany.
12Paper abstract
Advances in human pluripotent stem cell (hPSC) techniques have led them to become a widelyused and powerful tool for a vast array of applications, including disease modeling,developmental studies, drug discovery and testing, and emerging cell-based therapies. hPSC
workflows that require clonal expansion from single cells, such as CRISPR/Cas9-mediated genome
editing, face major challenges in terms of efficiency, cost, and precision. Classical subcloning
approaches depend on limiting dilution and manual colony picking, which are both timeconsuming and labor-intensive, and lack a real proof of clonality. Here we describe the applicationof three different automated cell isolation and dispensing devices that can enhance the singlecell cloning process for hPSCs. In combination with optimized cell culture conditions, these devices
offer an attractive alternative compared to manual methods. We explore various aspects of eachdevice system and define protocols for their practical application. Following the workflow
described here, single cell−derived hPSC sub-clones from each system maintain pluripotency andgenetic stability.
Technologies
In their paper the authors utilized three independent technologies to improve and further
automate their single-cell cloning workflows. These technologies were the isoCell (fromiotaSciences) and two commercially available single-cell dispensers. The authors state they
developed “a workflow in which the isoCell platform is applied to enable robust, efficient, costeffective, and easy-to-handle hPSC clone isolation and expansion”. The isoCell isthe only instrument to provide automation of all liquid-handling steps including single-cellisolation, media exchange and harvesting of resultant hPSC clonal cultures in sub-microliter
volumes. In contrast, both single-cell dispensers considered only performed the single-cellisolation part of the workflow.

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isoCell User Experience Summary
User experience
The authors considered three instruments for use in their hPSC cloning protocols. Based on these
protocols, a succinct overview of user experiences and some relevant instrument characteristics

are presented in Table 4 of their paper. This table is partially reproduced below in our comparison
table. The isoCell is the only solution that enables in-chamber verification of monoclonality

combined with start-to-finish liquid-handling automation for their complete single-cell cloning
workflows.

To find more information about the isoCell: www.iotasciences.com/isocell
Comparison Table
A partial reproduction of Table 4 from the paper provides a summary of the technologies
considered by the authors for their single-cell cloning protocols in the hPSC field.

iotaSciences

Aspect/Equipment

Single-cell

Single-cell

isoCelldispenser 1

dispenser 2

Training

Half-day3-5 days

1 dayHandlingEasy

DifficultModerate

Maintenance requirements

Low

Demanding

Low

Embedded in automated platform

NoYes

Yes

In-chamber
verification

Beforechamber
verification

Beforechamber
verification< 1 µl60 - 100 µl

60 - 100 µl

MediumHighHigh

Low

Low

High

User experience

Assurance of monoclonality
Proof of monoclonality
Cell culture volume

Cost
Hardware acquisition costs
Consumable cost

Access the open access journal paper here: https://doi.org/10.1002/cpsc.123

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