Microfluidic separation of axonal and somal compartments of neural progenitor cells differentiated in a 3D matrix

Madhura S. Lotlikar; Marina B. Tarantino; Mehdi Jorfi; Dora M. Kovacs; Rudolph E. Tanzi; Raja Bhattacharyya

STAR Protocols (Mar 2022)

Microfluidic separation of axonal and somal compartments of neural progenitor cells differentiated in a 3D matrix

Madhura S. Lotlikar,
Marina B. Tarantino,
Mehdi Jorfi,
Dora M. Kovacs,
Rudolph E. Tanzi,
Raja Bhattacharyya

Affiliations

Madhura S. Lotlikar: Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA
Marina B. Tarantino: Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA
Mehdi Jorfi: Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA
Dora M. Kovacs: Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA
Rudolph E. Tanzi: Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA
Raja Bhattacharyya: Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02115, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 1
p. 101028

Abstract

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Summary: This protocol describes the differentiation of human neural progenitor cells (hNPCs) in a microfluidic device containing a thin 3D matrix with two separate chambers, enabling a cleaner separation between axons and soma/bulk neurons. We have used this technique to study how mitochondria-associated ER membranes (MAMs) regulate the generation of somal and axonal amyloid β (Aβ) in FAD hNPCs, a cellular model of Alzheimer’s disease. This protocol also details the quantification of Aβ molecules and isolation of pure axons via axotomy.For complete details on the use and execution of this profile, please refer to Bhattacharyya et al. (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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