Application note

Cancer Research

Accelerating drug discovery (Data provided by Dr. Warren, UCSF)

The drug screening based on three-dimensional culture or Organoids is a state-of-the-art in precision medicine. We applied the YAMAHA CELL HANDLER™ to moderate throughput drug screening against patient-derived cancer spheroids.

Accurate 3D cell-based assay - targeted on ex vivo - (Collaboration with On-chip Biotechnologies Co., Ltd.)

3D cell-based assays using spheroids and ex vivo tumor fragments have recently been paid much attention in preclinical drug discovery and drug efficacy evaluation. However, size uniformity as well as cell viability need to be significantly improved for obtaining accurate and reproducible data. On-chip™ Sort (On-chip Biotechnologies Co., Ltd., Tokyo, Japan) can select intended samples from a population based on scattered light and fluorescence. YAMAHA CELL HANDLERTM (YCH) can select and transfer targeted samples from one plate to another. In this report, we describe a method for preparation of precise 3D cell-based assay by combining the two instruments.

Organoid Research

Direct picking from Matrigel^® dome (Collaboration with HUB) / Assay with two layer Matrigel^®

YAMAHA CELL HANDLER™ can directly and accurately pick up cell aggregates from hydrogel media, e.g. Matrigel^® (Figure 1 for two-layer Matrigel^® and Figure 2 for dome Matrigel^® ). This technology significantly shortens the duration of the spheroid or organoid collection from the MP and thereby increasing the assay throughput and testing of assay conditions.

Isolation of kidney organoids (Data provided by Dr. Takasato, RIKEN)

This laboratory at RIKEN is focused on uncovering the developmental mechanisms of human mesoderm in the kidney by utilizing its unique technology that can generate hiPSC-derived kidney organoids from pluripotent cells in vitro. The capability of the CELL HANDLER™ was evaluated in the selection and isolation of target kidney organoids exhibiting specific morphological characteristics.

Isolation of lung organoids (Data provided by Dr. Morimoto, RIKEN)

Research on the respiratory system at the RIKEN laboratory has focused on generating respiratory tissue organoids from mice. To develop an in vitro pathology model, uniform organoids are grown in semi-solid support such as Matrigel^®. The spheroids are selected and isolated based on morphological assessment, which is an important requirement in 3D culture. The applicability of CELL HANDLER™ in this spheroid transfer process was evaluated by the RIKEN research group.

Automated Spheroid Picking and Transfer from AggreWell™ Plate Using CELL HANDLER™2

The CELL HANDLER™2 ("YCH2") is a device that automatically images, sorts and moves target cells with high speed and precision. This application note presents the YCH2 performance evaluation of automated spheroid transfer from the AggreWell™ plate. The tests demonstrated efficient and effective selection and transfer of uniform spheroids by using the AI-based detection model and parameter-based filtering of target spheroids. The results showed that YCH2 moved 96 spheroids from each of the AggreWell™400 and AggreWell™800 plates in approximately 9 minutes, achieving a 99 % transfer success rate. The results suggest YCH2's usefulness in spheroid-based drug screening and applications in co-culture systems.

Automated Spheroid Picking and Transfer from EZSPHERE^® Plate Using CELL HANDLER™2

The CELL HANDLER™2 ("YCH2") is a device that automatically images, sorts and moves target cells with high speed and precision. This application note presents the YCH2 performance evaluation of automated spheroid transfer from the EZSPHERE^® SP microplate. The tests demonstrated efficient and effective selection and transfer of uniform spheroids by using the AI-based detection model and parameter-based filtering of target spheroids. The results showed that YCH2 moved 96 spheroids in approximately 10 minutes, achieving a 99 % cell transfer success rate. The results suggest YCH2's usefulness in spheroid-based drug screening and applications in co-culture systems.

Stem Cell Research

Automated iPS cell sorting by using CELL HANDLER™ (Data provided by Dr. Tsukahara and Dr. Ichisaka, CiRA Foundation)

The Facility for iPS Cell Therapy (FiT), as a part of the CiRA foundation, manufactures inducible pluripotent stem cells (iPS) for regenerative medicine research. In the manufacturing process, selection of high quality iPS cells plays an important role. This is accomplished entirely by manual operation, which requires an excessive amount of time and labor. CELL HANDLER™ is capable of imaging and recognizing cells one by one, in addition to accurately dispensing single cells and colonies. In this report, we describe an automated method for isolating single cells and colonies of iPS cells by using CELL HANDLER™.

Differentiation analysis of iPSC / iPS-derived cardiomyocytes (Data provided by Dr. Shimizu (Tokyo Women's Medical Univ., ABMES))

Embryoid body (EB) spheroids derived from 201B7 cells were selected and grouped by size (area). Expression levels of several differentiation markers were measured. Our result showed that small-sized EB spheroids tended to remain undifferentiated.

Single Cell Research

Development of a process to sort high antibody-producing cells using the CELL HANDLER™

The fast-growing antibody therapy market is experiencing the rapid development of new technologies. The isolation of high antibody-producing cells is critical and yet challenging, and the market requires improved and more efficient processes. The YAMAHA CELL HANDLER™ cell picking & imaging system automates the detection, isolation, and transfer of single cells. This cell isolation technology was used to develop a new process that sorts high antibody-producing cells in a highly efficient and effective manner.

Selection of transfectants / Imaging options for single cell

Establishing stable monoclonal transfectant cell lines requires drug selection followed by identification and isolation of single target cells (e.g., fluorescence-positive), clonal expansion, and verification—steps that can be labor-intensive when done manually. In this application note, CHO cells transfected with a GFP vector carrying a puromycin-resistance marker were selected with puromycin for 8 days, dispersed on an Elplasia^® plate, and single cells were detected by bright-field and fluorescence imaging. CELL HANDLER™ picked a fluorescent target cell and transferred it to another plate, forming a stably fluorescent colony after 6 days.

Establishment of clonal Organoid cultures after CRISPR-Cas9 genome editing using the Yamaha CELL HANDLER™ with HUB Organoids

Organoids are 3D tissues grown from stem cells and can be expanded long-term from many healthy or diseased samples. After CRISPR/Cas9 editing, cultures are typically polyclonal and editing efficiency is not 100%, so modified organoids (e.g., GFP-positive) must be identified and isolated. Because DNA repair can generate multiple genotypes, single-organoid picking, transfer to individual wells, expansion, and sequencing are needed to establish genetically pure clones—an increasingly labor-intensive workflow. The Yamaha CELL HANDLER™ aims to automate image-based recognition and transfer of target organoids using fluorescence and morphology, improving throughput and consistency in clonal selection.

Other applications

Automated placement of varied sized spheroids in Organ on a Chip, Mimetas OrganoPlate^® Graft by CELL HANDLER™

Advanced approach for 3D-cell phenotypic assay

Cell media change / Expanding cell culture / Picking of adherent spheroids

High-Accuracy Cell Detection Combining the CELL HANDLER™2 and the AI Image Recognition Model

CELL CELL HANDLER™2 performs image-based cell detection, analyzes and quantifies the features of detected cells, and transfers cells that meet specified parameters to an SBS-format culture plate in an automated workflow. Combining the instrument with an AI image recognition model enables stable, highly accurate detection across diverse cell types and culture plates, which expects to improve the operational efficiency of the CELL HANDLER™ even further. This application note presents the evaluation of cell detection accuracy using the AI image recognition model and its results.

High-Accuracy High-Efficiency Spheroid Transfer Using the CELL HANDLER™

Manual spheroid transfer is traditionally labor-intensive and prone to variability, creating bottlenecks in high-precision drug screening and applications involving Microphysiological System (MPS) plates. This study evaluates the performance of the CELL HANDLER™ for spheroid transfer compared to a conventional manual method. In the test, we used the CELL HANDLER™ to transfer spheroids derived from human colorectal cancer (HT-29 cell) and measured operation time, carried-over medium volume, cell transfer success rate, and morphology retention rate. Compared to the conventional manual method, the CELL HANDLER™ reduced operation time by approximately60 % and carried-over medium volume by approximately99 %, while achieving cell transfer success rates and morphology retention rates equal to or better than the manual method.

Accurate Automated Cell Transfer with Yamaha Precision Tip™ in Various Applications

Selecting the right tools for cell sizes and culture environment is essential in the automated cell transfer process. Yamaha Precision Tip™ (YPT) is a plastic tip with a syringe-like structure. With three size variations, YPT was developed exclusively to achieve highly efficient and accurate cell transfer via the CELL HANDLER™. The CELL HANDLER™ provides an automated cell handling workflow that includes attaching, calibrating, and disposing YPT, allowing accurate and reproducible cell manipulation. YPT may be used with not only single cells, but also suspended cells, 3D cells in gel culture media, and adherent 2D cells. This application note presents the cell transfer efficiency of the common CELL HANDLER™ use cases combined with YPT of different sizes.