Scalable generation of 3D pancreatic islet organoids from human pluripotent stem cells in suspension bioreactors

被引：3
|
作者：
Pollock, Samuel D. [1 ,2 ,3 ]
Galicia-Silva, Israeli M. [1 ,2 ,3 ]
Liu, Mai [2 ,3 ,4 ]
Gruskin, Zoe L. [1 ,2 ,3 ]
Alvarez-Dominguez, Juan R. [1 ,2 ,3 ]
机构：
[1] Univ Penn, Perelman Sch Med, Dept Cell & Dev Biol, Philadelphia, PA 19104 USA
[2] Univ Penn, Perelman Sch Med, Inst Regenerat Med, Philadelphia, PA 19104 USA
[3] Univ Penn, Perelman Sch Med, Inst Diabet Obes & Metab, Philadelphia, PA 19104 USA
[4] Univ Penn, Perelman Sch Med, Dept Bioengn, Philadelphia, PA 19104 USA
来源：
STAR PROTOCOLS | 2023年 / 4卷 / 04期
关键词：
MATURATION; REPLACEMENT; TRIGGERS;
D O I：
10.1016/j.xpro.2023.102580
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Here, we present a protocol for producing 3D pancreatic-like organoids from human pluripotent stem cells in suspension bioreactors. We describe scalable techniques for generating 10,000-100,000 organoids that further mature in 4- 5 weeks into a-and b -like cells with glucose-responsive insulin and glucagon release. We detail procedures for culturing, passaging, and cryopreserving stem cells as suspended clusters and specify growth media and differentiation factors for differentiation. Finally, we discuss functional assays for research appli-cations. For complete details on the use and execution of this protocol, please refer to Alvarez-Dominguez et al.1
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页数：40
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