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Worthington:Organoids 類器官,解離消化指南

發布者:艾美捷科技    發布時間:2026-01-28     
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類器官培養與功能研究,是酶解技術在發育生物學、腫瘤學、再生醫學及精準醫學等領域的核心應用方向。酶介導的組織消化與解離,作為類器官原代制備、體外擴增及細胞分離的關鍵步驟,已在各類器官源性類器官研究中被廣泛使用,但針對不同類器官的酶解作用適配機制、最佳解離條件的精準把控,仍為科研實踐中的重點難點。

 

研究人員在檢索類器官消化的適配酶種、最優濃度及配套培養體系的相關科學文獻時,往往會遇到實驗方案不統一、數據結果相互矛盾的問題。這種差異主要源于類器官的物種來源、組織源性差異,以及類器官自身細胞組成和基質結構的復雜性,同時也與酶制劑的選擇、使用濃度及解離操作條件的不同密切相關。

 

艾美捷匯總了 Worthington 針對不同物種、不同類型類器官的酶解消化方案,明確了適配的消化酶種類、使用濃度及配套的培養基體系,同時附列了對應的經典研究文獻參考。本類器官組織表為類器官的消化解離操作提供了標準化的實驗參考依據,為科研人員建立專屬的類器官酶解方案提供了合乎邏輯的實驗思路,并通過詳實的類器官特異性參考資料,助力開發穩定、有效的類器官消化解離方法。更多其他組織的解離指南,點擊查看:各組織分離指南。

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物種類器官 / 細胞類型消化酶及濃度培養基參考文獻
人類肺類器官1 型膠原酶:0.2%基礎培養基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
人類腦類器官PDS 試劑盒:詳見參考文獻磷酸鹽緩沖液(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
人類大腦類器官PDS 試劑盒:詳見參考文獻磷酸鹽緩沖液(PBS)Schr?ter, 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
人類(胚胎)肺類器官中性蛋白酶:24 u/mlDMEM/F12 培養基Nikoli?, 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
I 型脫氧核糖核酸酶:0.001%
人類結直腸癌類器官4 型膠原酶:0.25%DMEM/F-12 培養基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
I 型脫氧核糖核酸酶:0.01%
透明質酸酶:0.002%
人類視網膜類器官PDS 試劑盒:詳見參考文獻DMEM 培養基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
人類腦類器官木瓜蛋白酶:20 u/ml平衡鹽溶液(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
I 型脫氧核糖核酸酶:0.005%
人類大腦類器官木瓜蛋白酶:20 u/mlDMEM/F12 培養基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
I 型脫氧核糖核酸酶:0.01%
人類干細胞來源視網膜類器官PDS 試劑盒:詳見參考文獻平衡鹽溶液(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
人類腸道類器官4 型膠原酶:0.2%RPMI 培養基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
I 型脫氧核糖核酸酶:0.015%
小鼠(8-12 周)胰腺類器官中性蛋白酶:0.012%DMEM 培養基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
膠原酶:0.012%
小鼠結腸腫瘤類器官1 型膠原酶:0.2%磷酸鹽緩沖液(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
小鼠(胚胎)肺類器官中性蛋白酶:24 u/mlDMEM/F12 培養基Nikoli?, 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
I 型脫氧核糖核酸酶:0.001%
小鼠類器官培養物干細胞解離酶 1:0.1%磷酸鹽緩沖液(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
小鼠腸道類器官4 型膠原酶:500 u/ml基礎培養基O'Rourke, K., Ackerman, S., Dow, L. and Lowe, S. Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells. Bio Protoc 6, 2016
I 型脫氧核糖核酸酶:15 u/ml
小鼠結腸腫瘤類器官4 型膠原酶:200 u/mlDMEM 培養基Xue, X. and Shah, Y. In Vitro Organoid Culture of Primary Mouse Colon Tumors. J Vis Exp, e50210, 2013
中性蛋白酶:0.01%
小鼠腸道類器官1 型膠原酶:800 u/mlDMEM 培養基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
中性蛋白酶:0.013%
小鼠(成年)類器官4 型膠原酶:200-300 u/mlDMEM 培養基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
小鼠乳腺類器官1 型膠原酶:0.2%DMEM/F12 培養基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
胰蛋白酶:0.2%
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