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Optimized Protocol for Generating Functional Pancreatic Insulin-secreting Cells from Human Pluripotent Stem Cells
Optimized Protocol for Generating Functional Pancreatic Insulin-secreting Cells from Human Pluripotent Stem Cells
JoVE Journal
Biology
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JoVE Journal Biology
Optimized Protocol for Generating Functional Pancreatic Insulin-secreting Cells from Human Pluripotent Stem Cells

Optimized Protocol for Generating Functional Pancreatic Insulin-secreting Cells from Human Pluripotent Stem Cells

1,687 Views

06:33 min

February 02, 2024

DOI:

06:33 min
February 02, 2024

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Transcript

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Attaining a high and consistent efficiency in the differentiation process for converting pluripotent stem cells into human beta cells is a notable challenge. This challenge is influenced by the precision and the consistency with which the experimenter follows the protocol. Moreover, the amount of beta cells obtained in each experiment is quite variable, which may result in the reduced function of beta cells across multiple experiments.

In the context of high leptin beta cell research in diabetes, the availability of cadaveric eyelet donors is quite limited. Consequently, the use of stem cell differentiated beta cells is offering an unlimited and abundant supply of insulin secreting cells. Beta cell differentiation is a pivotal advancement in our field, offering numerous benefits.

It allows us to explore the development and function of beta cells, leading to a deeper understanding of diabetes causes and beta cell dysfunction. We aim to use this technique for our research focusing on the genetics of diabetes. Our primary objective is to focus on mutation of certain genes that can influence beta cells and eyelet function.

So this protocol will facilitate the access to human eyelet derived from pre-potent stem cells with mutations of interest.

Summary

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This article presents a protocol for directed differentiation and functional analysis of β-cell like cells. We describe optimal culture conditions and passages for human pluripotent stem cells before generating insulin-producing pancreatic cells. The six-stage differentiation progresses from definitive endoderm formation to functional β-cell like cells secreting insulin in response to glucose.

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