Organoids

Species	Cells	Enzyme(s)	Medium	Reference
Human Human	Lung Organoids	Collagenase Type 1 : 0.2%	Basal medium	Chiu, M. , Li, C. , Yu, Y. , Liu, X. , Huang, J. , Wan, Z. , Yuen, K. and Zhou, J. Establishing Bipotential Human Lung Organoid Culture System and Differentiation to Generate Mature Alveolar and Airway Organoids. , Bio Protoc 13 , e4657 , 2023
Human Human	Brain organoids	PDS kit : see reference	PBS	Velasco, S. , Kedaigle, A. , Simmons, S. , Nash, A. , Rocha, M. , Quadrato, G. , Paulsen, B. , Nguyen, L. , Adiconis, X. , Regev, A. , Levin, J. and Arlotta, P. Individual Brain Organoids Reproducibly Form Cell Diversity of the Human Cerebral Cortex. , Nature 570 , 523-527 , 2019
Human Human, adolescent	Epithelial organoids	Collagenase : 0.3% Neutral Protease : 0.4%	MEM	Hemeryck, L. , Hermans, F. , Chappell, J. , Kobayashi, H. , Lambrechts, D. , Lambrichts, I. , Bronckaers, A. and Vankelecom, H. Organoids from Human Tooth Showing Epithelial Stemness Phenotype and Differentiation Potential. , Cell Mol Life Sci 79 , 153 , 2022
Human Human	Cerebral organoids	PDS kit : see reference	PBS	Schröter, M. , Wang, C. , Terrigno, M. , Hornauer, P. , Huang, Z. , Jagasia, R. and Hierlemann, A. Functional Imaging of Brain Organoids Using High-Density Microelectrode Arrays. , MRS Bull Vol. 47 , 530-544 , 2022
Human Human	Lamina propria lymphocytes	Collagenase Type 4 : 0.1% Deoxyribonuclease I : 10 u/ml	DMEM/F-12	Schreurs, R. , Baumdick, M. , Drewniak, A. and Bunders, M. Co-culture of Human Intestinal Organoids and Lamina Propria-derived CD4 T Cells. , STAR Protoc Vol. 2 , 100519 , 2021
Human Human	Airway organoids	Pronase : 0.15% Collagenase : 0.05%	Airway medium	Vaart, J. , Böttinger, L. , Geurts, M. , Wetering, W. , Knoops, K. , Sachs, N. , Begthel, H. , Korving, J. , Lopez-Iglesias, C. , Peters, P. , Eitan, K. , Gileles-Hillel, A. and Clevers, H. Modelling of Primary Ciliary Dyskinesia using Patient-derived Airway Organoids. , EMBO Rep Vol. 22 , e52058 , 2021
Human Human, embryonic	Lung organoids	Neutral Protease : 24 u/ml Deoxyribonuclease I : 0.001%	DMEM/F12	Nikolić, M. , Caritg, O. , Jeng, Q. , Johnson, J. , Sun, D. , Howell, K. , Brady, J. , Laresgoiti, U. , Allen, G. , Butler, R. , Zilbauer, M. , Giangreco, A. and Rawlins, E. Human Embryonic Lung Epithelial Tips are Multipotent Progenitors that can be Expanded In vitro as Long-term Self-renewing Organoids. , Elife 6 , 2017
Human Human	Colorectal cancer organoids	Collagenase Type 4 : 0.25% Deoxyribonuclease I : 0.01% Hyaluronidase : 0.002%	DMEM/F-12	Marinucci, M. , Ercan, C. , Taha-Mehlitz, S. , Fourie, L. , Panebianco, F. , Bianco, G. , Gallon, J. , Staubli, S. , Soysal, S. , Zettl, A. , Rauthe, S. , Vosbeck, J. , Droeser, R. , Bolli, M. , Peterli, R. , Flüe, M. , Ng, C. , Kollmar, O. , Coto-Llerena, M. and Piscuoglio, S. Standardizing Patient-Derived Organoid Generation Workflow to Avoid Microbial Contamination From Colorectal Cancer Tissues. , Front Oncol Vol. 11 , 781833 , 2021
Human Human	Astrocytes	PDS kit : see reference	EBSS	Barbar, L. , Rusielewicz, T. , Zimmer, M. , Kalpana, K. and Fossati, V. Isolation of Human CD49f Astrocytes and iPSC-Based Neurotoxicity Assays. , STAR Protoc Vol. 1 , 100172 , 2020
Human Human	Retinal Organoids	PDS kit : see reference	DMEM	Tresenrider, A. , Sridhar, A. , Eldred, K. , Cuschieri, S. , Hoffer, D. , Trapnell, C. and Reh, T. Single-cell Sequencing of Individual Retinal Organoids Reveals Determinants of Cell Fate Heterogeneity. , bioRxiv , 2023
Human Human	Brain organoids	Papain : 20 u/ml Deoxyribonuclease I : 0.005%	EBSS	Lisowski, P. , Lickfett, S. , Rybak-Wolf, A. , Menacho, C. and Le, S. Mutant Huntingtin Impairs Neurodevelopment in Human Brain Organoids through CHCHD2-mediated Neurometabolic Failure. , Nat Commun 15 , 7027 , 2024
Human Human	Cerebral organoids	Papain : 20 u/ml Deoxyribonuclease I : 0.01%	DMEM/F12	Geest, A. , Jakobs, C. , Ljubikj, T. , Huffels, C. , Luna, M. , Sá, R. , Adolfs, Y. , Wit, M. , Rutten, D. , Kaal, M. , Zwartkruis, M. , Carcolé, M. , Groen, E. , Hol, E. , Basak, O. , Isaacs, A. , Westeneng, H. , Berg, L. , Veldink, J. , Schlegel, D. and Pasterkamp, R. Molecular Pathology, Developmental Changes, and Synaptic Dysfunction in (pre-) Symptomatic Human C9ORF72-ALS/FTD Cerebral Organoids. , Acta Neuropathol Commun Vol. 12 , 152 , 2024
Human Human	Stem cell-derived retinal organoids	PDS kit : see reference	EBSS	Xue, Y. , Seiler, M. , Tang, W. , Wang, J. , Delgado, J. , McLelland, B. , Nistor, G. , Keirstead, H. and Browne, A. Retinal Organoids On-a-Chip: a Micro-Millifluidic Bioreactor for Long-term Organoid Maintenance. , Lab Chip 21 , 3361-3377 , 2021
Human Human	Intestinal organoids	Collagenase Type 4 : 0.2% Deoxyribonuclease I : 0.015%	RPMI	Gamero-Estevez, E. , Hensel, I. , Steinhauer, M. , Müllertz, O. , Savochkina, E. , Sektioglu, I. , Stoll, B. , Derakhshani, S. , Devriese, S. , Kim, K. and Resnik-Docampo, M. Protocol for Immune Cell Isolation, Organoid Generation, and Co-culture Establishment from Cryopreserved Whole Human Intestine. , Bio Protoc 15 , e5157 , 2025
Mouse Mouse, 8-12 week	Pancreas organoid	Neutral Protease : 0.012% Collagenase : 0.012%	DMEM	Huch, M. , Bonfanti, P. , Boj, S. , Sato, T. , Loomans, C. , Van de Wetering, M. , Sojoodi, M. , Li, V. , Schuijers, J. , Gracanin, A. , Ringnalda, F. , Begthel, H. , Hamer, K. , Mulder, J. , Van Es, J. and De Koning, E. Unlimited In Vitro Expansion of Adult Bi-Potent Pancreas Progenitors Through the Lgr5/R-Spondin Axis. , EMBO J 32 , 2708-21 , 2013
Mouse Mouse	Colonic tumor organoids	Collagenase Type 1 : 0.2%	PBS	Klemke, L. , Blume, J. , Oliveira, T. and Schulz-Heddergott, R. Preparation and Cultivation of Colonic and Small Intestinal Murine Organoids Including Analysis of Gene Expression and Organoid Viability. , Bio Protoc 12 , e4298 , 2022
Mouse Mouse, embryonic	Lung organoids	Neutral Protease : 24 u/ml Deoxyribonuclease I : 0.001%	DMEM/F12	Nikolić, M. , Caritg, O. , Jeng, Q. , Johnson, J. , Sun, D. , Howell, K. , Brady, J. , Laresgoiti, U. , Allen, G. , Butler, R. , Zilbauer, M. , Giangreco, A. and Rawlins, E. Human Embryonic Lung Epithelial Tips are Multipotent Progenitors that can be Expanded In vitro as Long-term Self-renewing Organoids. , Elife 6 , 2017
Mouse Mouse	Organoid cultures	Stemxyme 1 : 0.1%	PBS	Huang, L. , Desai, R. , Conrad, D. , Leite, N. , Akshinthala, D. , Lim, C. , Gonzalez, R. , Muthuswamy, L. , Gartner, Z. and Muthuswamy, S. Commitment and Oncogene-induced Plasticity of Human Stem Cell-derived Pancreatic Acinar and Ductal Organoids. , Cell Stem Cell Vol. 28 , 1090-1104.e6 , 2021
Mouse Mouse	Lung	Collagenase Type 1 : 450 u/ml Elastase : 1 u/ml Neutral Protease : 5 u/ml Deoxyribonuclease I : 0.005%	DMEM/F12	Fujimura, T. , Enomoto, Y. , Katsura, H. , Ogawa, T. , Baba, S. , Ogata, A. , Yamaoka, A. , Shiroguchi, K. and Morimoto, M. Identifying a Lung Stem Cell Subpopulation by Combining Single-Cell Morphometrics, Organoid Culture, and Transcriptomics. , Stem Cells 41 , 809-820 , 2023
Mouse Mouse	Intestinal organoids	Collagenase Type 4 : 500 u/ml Deoxyribonuclease I : 15 u/ml	Basal Medium	O'Rourke, K. , Ackerman, S. , Dow, L. and Lowe, S. Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells. , Bio Protoc 6 , 2016
Mouse Mouse	Colon tumor organoids	Collagenase Type 4 : 200 u/ml Neutral Protease : 0.01%	DMEM	Xue, X. and Shah, Y. In Vitro Organoid Culture of Primary Mouse Colon Tumors. , J Vis Exp , e50210 , 2013
Mouse Mouse	Intestinal organoids	Collagenase Type 1 : 800 u/ml Neutral Protease : 0.013%	DMEM	Barthel, E. , Speer, A. , Levin, D. , Sala, F. , Hou, X. , Torashima, Y. , Wigfall, C. and Grikscheit, T. Tissue Engineering of the Intestine in a Murine Model. , J Vis Exp 70 , e4279 , 2012
Mouse Mouse, adult	Organoids	Collagenase Type 4 : 200-300 u/ml	DMEM	Neal, J,, Li ,X. , Zhu, J. , Giangarra, V. , Grzeskowiak, C. , Ju, J. , Liu, I. Chiou, S. , Salahudeen, A. and Smith, A. et al Organoid Modeling of the Tumor Immune Microenvironment. , Cell 175 , 1972 , 2018
Mouse Mouse	Mammary organoids	Collagenase Type 1 : 0.2% Trypsin : 0.2%	DMEM/F12	Charifou, E. , Sumbal, J. , Koledova, Z. , Li, H. and Chiche, A. A Robust Mammary Organoid System to Model Lactation and Involution-like Processes. , Bio Protoc 11 , e3996 , 2021