Massively Parallel High-Throughput Single-Cell Patterning and Large Biomolecular Delivery in Mammalian Cells Using Light Pulses.

cell viability infrared light pulses large biomolecular delivery microcontact printing parallel single-cell patterning transfection efficiency

Journal

Small (Weinheim an der Bergstrasse, Germany)
ISSN: 1613-6829
Titre abrégé: Small
Pays: Germany
ID NLM: 101235338

Informations de publication

Date de publication:
Nov 2023
Historique:
revised: 21 06 2023
received: 11 04 2023
medline: 23 11 2023
pubmed: 7 8 2023
entrez: 7 8 2023
Statut: ppublish

Résumé

The recent advancements of single-cell analysis have significantly enhanced the ability to understand cellular physiology when compared to bulk cellular analysis. Here a massively parallel single-cell patterning and very large biomolecular delivery is reported. Micro-pillar polydimethyl siloxane stamp with different diameters (40-100 µm with 1 cm × 1 cm patterning area) is fabricated and then imprint distinct proteins and finally pattern single-cell to small clusters of cells depending on the micro-pillar diameters. The maximum patterning efficiency is achieved 99.7% for SiHa, 96.75% for L929, and 98.6% for MG63 cells, for the 100 µm micro-pillar stamp. For intracellular delivery of biomolecules into the patterned cells, a titanium micro-dish device is aligned on top of the cells and exposed by infrared light pulses. The platform successfully delivers small to very large biomolecules such as PI dyes (668 Da), dextran 3000 Da, siRNA (20-24 bp), and large size enzymes (464 KDa) in SiHa, L929 and MG63 cells. The delivery efficiency for PI dye, Dextran 3000, siRNA, and enzyme for patterned cells are ≈95 ± 3%, 97 ± 1%, 96 ± 1% and 94 ± 3%, with cell viability of 98 ± 1%. Thus, the platform is compact, robust, easy for printing, and potentially applicable for single-cell therapy and diagnostics.

Identifiants

pubmed: 37548122
doi: 10.1002/smll.202303053
doi:

Substances chimiques

Dextrans 0
Proteins 0
RNA, Small Interfering 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

e2303053

Subventions

Organisme : DBT/Wellcome Trust India Alliance Fellowship
ID : IA/E/16/1/503062

Informations de copyright

© 2023 Wiley-VCH GmbH.

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Auteurs

Gayathri R (G)

Department of Mechanical Engineering, Indian Institute of Technology Madras, Chennai, 600036, India.
Department of Engineering Design, Indian Institute of Technology Madras, Chennai, 600036, India.

Srabani Kar (S)

Department of Engineering Design, Indian Institute of Technology Madras, Chennai, 600036, India.
Department of Physics, Indian Institute of Science Education and Research, Tirupati, 517507, India.

Moeto Nagai (M)

Department of Mechanical Engineering, Toyohashi University of Technology, Aichi, 441-8580, Japan.

Pallab Sinha Mahapatra (PS)

Department of Mechanical Engineering, Indian Institute of Technology Madras, Chennai, 600036, India.

Tuhin Subhra Santra (TS)

Department of Engineering Design, Indian Institute of Technology Madras, Chennai, 600036, India.

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