Unique single-use culture vessel

Rotating the cylindrical culture vessel with syringe function realizes 3D rotational floating culture and facilitates sampling of cells and medium after culture.
The culture environment of the externally sealed culture vessel is extremely low risk for fungal and mold contamination, and the air-permeable resin provides a culture environment similar to an air environment.

Cultured cell spheroids

The medium flow generated by slow rotation exerts a constant shear stress load on the cells, and the cultured spheroids tend to be spherical in shape and generally uniform in size.

Examples of culture results

iPS cells, ES cells, MSC cells, differentiated cells derived from iPS cells, human-derived cancer cells, etc.

Growth images of iPS cell spheroids

NEW

Chondrogenic differentiation from hMSC spheroids

We generated spheroids of human mesenchymal stem cells using CellPet 3D-iPS and induced chondrogenic differentiation by switching the culture medium. Compared to chondrogenic differentiation in monolayer culture, the spheroids produced greater amounts of hyaluronic acid and elastic fibers. Additionally, cartilage lumen, a characteristic feature of mature cartilage tissue, were observed within the tissue.

NEW

Gas permeability evaluation of disposable culture vessels

We evaluated the gas permeability of the disposable vessel of CellPet 3D-iPS. After removing oxygen and carbon dioxide from the culture medium by nitrogen bubbling, the extent to which each gas permeated the vessel and dissolved into the culture medium was examined. After about one hour, the concentration of dissolved oxygen in the vessels reached the same level as that in the culture dish.

hMSC spheroid culture using CellPet 3D-iPS

We performed spheroid culture of human mesenchymal stem cells using CellPet 3D-iPS. The cells were seeded into a culture vessel, and the medium was replaced every three days. On day six, the spheroids were enzymatically dissociated into single cells, and the cells were divided into two equal parts, which were then seeded into two separate culture vessels to continue the culture. Over the 12-day culture period, we observed an approximately eight-fold increase in cell proliferation. The diameter distribution of the obtained spheroids was uniform, and it was confirmed that the characteristics of mesenchymal stem cells were maintained.

iPS spheroid culture using CellPet 3D-iPS

We performed spheroid culture of iPS cells using CellPet 3D-iPS. Upon evaluating the size distribution of the spheroids on day 4 of culture, approximately 60% had a diameter of 200-300 μm, indicating the formation of uniform iPS spheroids. It was also confirmed that the expression of undifferentiated markers was maintained during culture with CellPet 3D-iPS.

Gas-permeable ability of disposable culture vessels

We performed spheroid cultures of iPS cells and osteosarcoma cells using both a newly developed disposable culture vessel and a conventional glass culture vessel. The disposable vessel, which has excellent gas permeability, was able to maintain a high dissolved oxygen concentration during spheroid culture. Additionally, the use of the disposable vessel enabled the formation of spheroids in osteosarcoma cells, which was not possible with the glass vessel.

iPS trilineage differentiation using CellPet 3D-iPS

We performed 3D rotary floating cultures of iPS cells using CellPet 3D-iPS. When using APEL2 medium, an increase in the expression of the trilineage differentiation markers Sox1, PDGFRα, and Sox17 was observed. This suggests that spheroid cultures using APEL2 medium and CellPet 3D-iPS can induce differentiation.