Protocol for fabricating elastomeric stencils for patterned stem cell differentiation

Summary: Geometrically controlled stem cell differentiation promotes reproducible pattern formation. Here, we present a protocol to fabricate elastomeric stencils for patterned stem cell differentiation. We describe procedures for using photolithography to produce molds, followed by molding polydime...

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Main Authors: Stefanie Lehr, Jack Merrin, Monika Kulig, Thomas George Minchington, Anna Kicheva
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724003526
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author Stefanie Lehr
Jack Merrin
Monika Kulig
Thomas George Minchington
Anna Kicheva
author_facet Stefanie Lehr
Jack Merrin
Monika Kulig
Thomas George Minchington
Anna Kicheva
author_sort Stefanie Lehr
collection DOAJ
description Summary: Geometrically controlled stem cell differentiation promotes reproducible pattern formation. Here, we present a protocol to fabricate elastomeric stencils for patterned stem cell differentiation. We describe procedures for using photolithography to produce molds, followed by molding polydimethylsiloxane (PDMS) to obtain stencils with through holes. We then provide instructions for culturing cells on stencils and, finally, removing stencils to allow colony growth and cell migration. This approach yields reproducible two-dimensional organoids tailored for quantitative studies of growth and pattern formation.For complete details on the use and execution of this protocol, please refer to Lehr et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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institution Kabale University
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publishDate 2024-12-01
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series STAR Protocols
spelling doaj-art-c34e49a81b8b452f94d1a29d8e6a8e472024-11-27T05:03:11ZengElsevierSTAR Protocols2666-16672024-12-0154103187Protocol for fabricating elastomeric stencils for patterned stem cell differentiationStefanie Lehr0Jack Merrin1Monika Kulig2Thomas George Minchington3Anna Kicheva4Institute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, AustriaInstitute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, AustriaInstitute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, AustriaInstitute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, AustriaInstitute of Science and Technology Austria, Am Campus 1, 3400 Klosterneuburg, Austria; Corresponding authorSummary: Geometrically controlled stem cell differentiation promotes reproducible pattern formation. Here, we present a protocol to fabricate elastomeric stencils for patterned stem cell differentiation. We describe procedures for using photolithography to produce molds, followed by molding polydimethylsiloxane (PDMS) to obtain stencils with through holes. We then provide instructions for culturing cells on stencils and, finally, removing stencils to allow colony growth and cell migration. This approach yields reproducible two-dimensional organoids tailored for quantitative studies of growth and pattern formation.For complete details on the use and execution of this protocol, please refer to Lehr et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724003526Developmental biologyCell DifferentiationOrganoidsBiotechnology and bioengineering
spellingShingle Stefanie Lehr
Jack Merrin
Monika Kulig
Thomas George Minchington
Anna Kicheva
Protocol for fabricating elastomeric stencils for patterned stem cell differentiation
STAR Protocols
Developmental biology
Cell Differentiation
Organoids
Biotechnology and bioengineering
title Protocol for fabricating elastomeric stencils for patterned stem cell differentiation
title_full Protocol for fabricating elastomeric stencils for patterned stem cell differentiation
title_fullStr Protocol for fabricating elastomeric stencils for patterned stem cell differentiation
title_full_unstemmed Protocol for fabricating elastomeric stencils for patterned stem cell differentiation
title_short Protocol for fabricating elastomeric stencils for patterned stem cell differentiation
title_sort protocol for fabricating elastomeric stencils for patterned stem cell differentiation
topic Developmental biology
Cell Differentiation
Organoids
Biotechnology and bioengineering
url http://www.sciencedirect.com/science/article/pii/S2666166724003526
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