A Simplified Method for Generating Kidney Organoids from Human Pluripotent Stem Cells
Kidney organoids generated from hPSCs have provided an unlimited source of renal tissue. Human kidney organoids are an invaluable tool for studying kidney disease and injury, developing cell-based therapies, and testing new therapeutics. For such applications, large numbers of uniform organoids and...
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| Veröffentlicht in: | Journal of Visualized Experiments 2021-04 (170) |
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| creator | Przepiorski, Aneta Crunk, Amanda E. Holm, Teresa M. Sander, Veronika Davidson, Alan J. Hukriede, Neil A. |
| description | Kidney organoids generated from hPSCs have provided an unlimited source of renal tissue. Human kidney organoids are an invaluable tool for studying kidney disease and injury, developing cell-based therapies, and testing new therapeutics. For such applications, large numbers of uniform organoids and highly reproducible assays are needed. We have built upon our previously published kidney organoid protocol to improve the overall health of the organoids. This simple, robust 3D protocol involves the formation of uniform embryoid bodies in minimum component medium containing lipids, insulin-transferrin-selenium-ethanolamine supplement and polyvinyl alcohol with GSK3 inhibitor (CHIR99021) for 3 days, followed by culture in knock-out serum replacement (KOSR)-containing medium. In addition, agitating assays allows for reduction in clumping of the embryoid bodies and maintaining a uniform size, which is important for reducing variability between organoids. Overall, the protocol provides a fast, efficient, and cost-effective method for generating large quantities of kidney organoids. |
| doi_str_mv | 10.3791/62452 |
| format | Article |
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Human kidney organoids are an invaluable tool for studying kidney disease and injury, developing cell-based therapies, and testing new therapeutics. For such applications, large numbers of uniform organoids and highly reproducible assays are needed. We have built upon our previously published kidney organoid protocol to improve the overall health of the organoids. This simple, robust 3D protocol involves the formation of uniform embryoid bodies in minimum component medium containing lipids, insulin-transferrin-selenium-ethanolamine supplement and polyvinyl alcohol with GSK3 inhibitor (CHIR99021) for 3 days, followed by culture in knock-out serum replacement (KOSR)-containing medium. In addition, agitating assays allows for reduction in clumping of the embryoid bodies and maintaining a uniform size, which is important for reducing variability between organoids. Overall, the protocol provides a fast, efficient, and cost-effective method for generating large quantities of kidney organoids.</description><subject>blood serum</subject><subject>Cell Culture Techniques - methods</subject><subject>Cell Differentiation</subject><subject>cost effectiveness</subject><subject>Developmental Biology</subject><subject>Humans</subject><subject>Induced Pluripotent Stem Cells - metabolism</subject><subject>Kidney - physiopathology</subject><subject>kidney diseases</subject><subject>kidneys</subject><subject>organoids</subject><subject>Organoids - metabolism</subject><subject>polyvinyl alcohol</subject><subject>therapeutics</subject><issn>1940-087X</issn><issn>1940-087X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV9LwzAUxYMo_pl-AR8kL4Iv1aRp2uZFkKGbqEyYA99C2t7OjDaZSTvYtze6Kfok9yEX8svh5ByETii5ZJmgV2mc8HgHHVKRkIjk2evur_0AHXm_ICSNCc_30QFjguW5iA_R7AZPdbtsdK2hwk_QvdkK19bhERhwqtNmjh90ZWCNJ26ujNWVx7WzLR73rTL4uemdXtoOTIenHbR4CE3jj9FerRoPJ9tzgGZ3ty_DcfQ4Gd0Pbx6jMqG8i6DkIuFUFVTx4KzMmKIqI5wQzrIy5axiJKtEEgOURUUzLmrCgBWp4qQogLABut7oLvuihaoMLpxq5NLpVrm1tErLvzdGv8m5XUkRsmAsDQIXWwFn33vwnWy1L8MXlAHbexlnNI8553n-P8pjmogwSUDPN2jprPcO6h9HlMjPtuRXW4E7-23_h_quJwCnG2BhVyAXtncmxLl9_QGG0Zet</recordid><startdate>20210413</startdate><enddate>20210413</enddate><creator>Przepiorski, Aneta</creator><creator>Crunk, Amanda E.</creator><creator>Holm, Teresa M.</creator><creator>Sander, Veronika</creator><creator>Davidson, Alan J.</creator><creator>Hukriede, Neil A.</creator><general>MyJove Corporation</general><scope>BEELZ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20210413</creationdate><title>A Simplified Method for Generating Kidney Organoids from Human Pluripotent Stem Cells</title><author>Przepiorski, Aneta ; Crunk, Amanda E. ; Holm, Teresa M. ; Sander, Veronika ; Davidson, Alan J. ; Hukriede, Neil A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c415t-ec59451ab1a5205c73a1a70500537c653d307d942eecbd1759f03e3b6a50bbe03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>blood serum</topic><topic>Cell Culture Techniques - methods</topic><topic>Cell Differentiation</topic><topic>cost effectiveness</topic><topic>Developmental Biology</topic><topic>Humans</topic><topic>Induced Pluripotent Stem Cells - metabolism</topic><topic>Kidney - physiopathology</topic><topic>kidney diseases</topic><topic>kidneys</topic><topic>organoids</topic><topic>Organoids - metabolism</topic><topic>polyvinyl alcohol</topic><topic>therapeutics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Przepiorski, Aneta</creatorcontrib><creatorcontrib>Crunk, Amanda E.</creatorcontrib><creatorcontrib>Holm, Teresa M.</creatorcontrib><creatorcontrib>Sander, Veronika</creatorcontrib><creatorcontrib>Davidson, Alan J.</creatorcontrib><creatorcontrib>Hukriede, Neil A.</creatorcontrib><collection>JoVE Journal: Developmental Biology</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Visualized Experiments</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Przepiorski, Aneta</au><au>Crunk, Amanda E.</au><au>Holm, Teresa M.</au><au>Sander, Veronika</au><au>Davidson, Alan J.</au><au>Hukriede, Neil A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Simplified Method for Generating Kidney Organoids from Human Pluripotent Stem Cells</atitle><jtitle>Journal of Visualized Experiments</jtitle><addtitle>J Vis Exp</addtitle><date>2021-04-13</date><risdate>2021</risdate><issue>170</issue><issn>1940-087X</issn><eissn>1940-087X</eissn><abstract>Kidney organoids generated from hPSCs have provided an unlimited source of renal tissue. 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| subjects | blood serum Cell Culture Techniques - methods Cell Differentiation cost effectiveness Developmental Biology Humans Induced Pluripotent Stem Cells - metabolism Kidney - physiopathology kidney diseases kidneys organoids Organoids - metabolism polyvinyl alcohol therapeutics |
| title | A Simplified Method for Generating Kidney Organoids from Human Pluripotent Stem Cells |
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