Isolation of lung organoids (Data provided by Dr. Morimoto, RIKEN)
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Laboratory for Lung Development and Regeneration
RIKEN Center for Biosystems Dynamics Research
Team Leader: Mitsuru Morimoto, Ph.D. / Yasunori Enomoto, M.D., Ph.D.
Background/Objective
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.
Material and Methods
Respiratory tissue organoids were generated in 75% Matrigel® domes in 24-well cell culture plates, subjected to morphological imaging and subsequently harvested directly from the domes using the CELL HANDLER™.
Observations obtained
Organoids which met pre-defined morphological criteria were identified (Fig. 1A), and were picked without contamination with adjacent organoids (Fig. 1B). A representative image of a single organoid transferred to a 96-well cell culture plate is shown in Fig. 2. These data confirmed that only selected organoids were reliably isolated by the CELL HANDLER™. In addition, growth of isolated organoids was observed (data not shown), indicating that no damage was sustained in the transfer process. These data obtained by morphology-based, direct harvesting of organoids from Matrigel® domes demonstrate the applicability of the CELL HANDLER™ in respiratory tissue organoid research.
Fig. 1. Images of organoids generated in a Matrigel® dome before (A) and after (B) picking by the CELL HANDLER™. Position of the organoid of interest is indicated by an arrow.
Fig. 2. Image of the organoid harvested.
Acknowledgement
Yamaha Motor would like to express its sincere thanks to Dr. M. Morimoto and Dr. Y. Enomoto for their guidance, experimental verification of the CELL HANDLER™ and for data they provided.
* "Matrigel" is a registered trademark of Corning Inc..
* For research use only. Not for use in diagnostic or therapeutic procedures.
* The specifications are subject to change without notice.
* The above are the results of experiments in our laboratory. The results may vary depending on the work environment, cell type and so on.
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