General Protocols
- Spheroid Culture on Pillar Plate
- Cell Suspension Culture in Matrigel on Pillar Plate
- Cell Suspension Culture in Alginate on Pillar Plate
- Dynamic Cell Culture in Perfusion Plate
- Dynamic Cell Culture with PetriLid
- Cell-Based Assays on Pillar Plate
- Liver Organoid Culture by 3D Bioprinting
- Cerebral Organoid Culture by Spheroid Transfer
- Troubleshooting Guide
Video Clips
Overview of cell loading and culture methods
Dynamic and static organoid cultures in the petri dish
Static organoid culture in the deep well plate
Dynamic organoid culture in the perfusion plate
Manual dispensing of cells in Matrigel on the pillar plate
Manual loading of cells in Matrigel by pillar stamping
Manual loading of cells in alginate by pillar stamping
Immunofluorescence staining of organoids on the pillar plate
Bioprinting of cells suspended in Matrigel on the pillar plate
Posters
Publications
- Lekkala, V.K.R., Shrestha, S., Qaryoute, A.A., Dhinoja, S., Acharya, P., Raheem, A., Jagadeeswaran, P., and Lee, M.Y., Enhanced maturity and functionality of vascularized human liver organoids through 3D bioprinting and pillar plate culture, BioRxiv, doi.org/10.1101/2024.08.21.608997 (2024)
Model: Vascularized human liver organoids
Application: Hepatotoxicity assays
- Zolfaghar, M., Acharya, P., Joshi, P., Choi, N.Y., Shrestha, S., Lekkala, V.K.R., Kang, S.Y., Lee, M.S., and Lee, M.Y., Cryopreservation of neuroectoderm on a pillar plate and in situ differentiation into human brain organoids, ACS Biomaterials Science & Engineering, doi.org/10.1021/acsbiomaterials.4c01383 (2024)
Model: Brain organoids
Application: On-chip cryopreservation of organoids
- Shrestha, S., Acharya, P., Kang, S.Y., Vanga, M.G., Lekkala, V.K.R., Liu, J., Yang, Y., Joshi, P., and Lee, M.Y Regenerative human liver organoids (HLOs) in a pillar/perfusion plate for hepatotoxicity assays, BioRxiv, doi.org/10.1101/2024.03.25.586638 (2024)
Model: Liver organoids
Application: Liver organoid regeneration and hepatotoxicity assays
- Acharya, P., Shrestha, S., Joshi, P., Choi, N.Y., Lekkala, V.K.R., Kang, S.Y., Ni, G., and Lee, M.Y., Dynamic culture of cerebral organoids using a pillar/perfusion plate for the assessment of developmental neurotoxicity, Biofabrication, doi.org/10.1088/1758-5090/ad867e (2024)
Model: Brain organoids
Application: Developmental neurotoxicity
- Lekkala, V.K.R., Kang, S.Y., Liu, J., Shrestha, S., Acharya, P., Joshi, P., Zolfaghar, M., Lee, M.S., Vanga, M.G., Jamdagneya, P., Pagnis, S., Kundi, A., Kabbur, S., Kim, U.T., Yang, Y., and Lee, M.Y., A pillar/perfusion plate enhances cell growth, reproducibility, throughput, and user friendliness in dynamic 3D cell culture, ACS Biomaterials Science & Engineering, doi.org/10.1021/acsbiomaterials.4c00179 (2024)
Model: 3D cell culture
Application: Dynamic spheroid culture for drug testing
- Shrestha, S., Lekkala, V.K.R., Acharya, P., Kang, S.Y., Vanga, M.G., and Lee, M.Y., Reproducible generation of human liver organoids (HLOs) on a pillar plate platform via microarray 3D bioprinting, Lab on a chip, DOI: 10.1039/D4LC00149D (2024)
Model: Liver organoid
Application: Hepatotoxicity
- Acharya, P., Joshi, P., Shrestha, S., Choi, N.Y., Jeong, S.H., and Lee, M.Y., Uniform cerebral organoid culture on a pillar plate by simple and reproducible spheroid transfer from an ultralow attachment well plate, Biofabrication, doi.org/10.1088/1758-5090/ad1b1e (2024)
Model: Brain organoid
Application: Developmental neurotoxicity
- Kang, S.Y., Kimura, M., Shrestha, S., Lewis, P., Lee, S.J., Cai, Y., Joshi, P., Acharya, P., Liu, J., Yang, Y., Sanchez, J.G., Ayyagari, S., Alsberg, E., Wells, J.M., Takebe, T., and Lee, M.Y., A pillar and perfusion plate platform for robust human organoid culture and analysis, Advanced Healthcare Materials, DOI: 10.1002/adhm.202302502 (2023)
Model: Liver and intestine organoids
Application: Pillar/perfusion plate for disease modeling
- Joshi, P., Kang, S.Y., Acharya, P., Sidhpura, D., and Lee, M.Y., High-throughput assessment of metabolism-mediated neurotoxicity by combining 3D-cultured neural stem cells and liver cell spheroids, Toxicology In Vitro, DOI: 10.1016/j.tiv.2023.105688 (2023)
Model: Liver and brain co-culture
Application: Metabolism-induced neurotoxicity
- Joshi, P., Patel, R., Kang, S.Y., Serbinowski, E., and Lee, M.Y., Establishment of ion channel and ABC transporter assays in 3D-cultrued ReNcell VM on a 384-pillar plate for neurotoxicity potential, Toxicology In Vitro, DOI: 10.1016/j.tiv.2022.105375 (2022)
Model: 3D neural stem cell culture
Application: Neurotoxicity
- Kang, S.Y., Joshi, P., and Lee, M.Y., High-throughput screening of compound neurotoxicity using 3D-cultured neural stem cells on a 384-pillar plate, Current Protocols in Toxicology, doi.org/10.1002/cpz1.107 (2021)
Model: 3D neural stem cell culture
Application: High-throughput screening
- Bigdelou, P., Chan, K.K., Tang, J.S., Yu, K.N., Whited, J., Wang, D., Lee, M.Y., and Sun, X.L., High-throughput multiplex assays with mouse macrophages on pillar plate platforms, Experimental Cell Research, 396, 112243, doi.org/10.1016/j.yexcr.2020.112243 (2020)
Model: Mouse macrophages
Application: High-throughput immunoassays
- Joshi, P., Kang, S.Y., Yu, K.N., Kothapalli, C., and Lee, M.Y., High-content imaging of 3D-cultured neural stem cells on a 384-pillar plate for the assessment of cytotoxicity, Toxicology In Vitro, 65, 104765, DOI: 10.1016/j.tiv.2020.104765 (2020)
Model: 3D cell culture
Application: Neurotoxicity
- Yu, K.N., Kang, S.Y., Hong, S., and Lee, M.Y., High-throughput metabolism-induced toxicity assays demonstrated on a 384-pillar plate, Archives of Toxicology, 92, 2501-2516, DOI: 10.1007/s00204-018-2249-1 (2018)
Model: 3D bioprinting
Application: Metabolism-induced hepatotoxicity
