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Single cell-laden protease-sensitive microniches for long-term culture in 3D.
Lienemann, Philipp S; Rossow, Torsten; Mao, Angelo S; Vallmajo-Martin, Queralt; Ehrbar, Martin; Mooney, David J.
Afiliação
  • Lienemann PS; John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA. mooneyd@seas.harvard.edu and Wyss Institute for Biologically Inspired Engineering, Cambridge, MA 02138, USA.
  • Rossow T; John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA. mooneyd@seas.harvard.edu and Wyss Institute for Biologically Inspired Engineering, Cambridge, MA 02138, USA.
  • Mao AS; John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA. mooneyd@seas.harvard.edu and Wyss Institute for Biologically Inspired Engineering, Cambridge, MA 02138, USA.
  • Vallmajo-Martin Q; Department of Obstetrics, University Hospital Zurich, University of Zurich, Schmelzbergstr. 12, 8091 Zurich, Switzerland.
  • Ehrbar M; Department of Obstetrics, University Hospital Zurich, University of Zurich, Schmelzbergstr. 12, 8091 Zurich, Switzerland.
  • Mooney DJ; John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA. mooneyd@seas.harvard.edu and Wyss Institute for Biologically Inspired Engineering, Cambridge, MA 02138, USA.
Lab Chip ; 17(4): 727-737, 2017 02 14.
Article em En | MEDLINE | ID: mdl-28154867
ABSTRACT
Single cell-laden three-dimensional (3D) microgels that can serve to mimic stem cell niches in vitro, and are therefore termed microniches, can be efficiently fabricated by droplet-based microfluidics. In this technique an aqueous polymer solution along with a highly diluted cell solution is injected into a microfluidic device to create monodisperse pre-microgel droplets that are then solidified by a polymer crosslinking reaction to obtain monodisperse single cell-laden microniches. However, problems limiting this approach studying the fate of single cells include Poisson encapsulation statistics that result in mostly empty microniches, and cells egressing from the microniches during subsequent cell culture. Here, we present a strategy to bypass Poisson encapsulation statistics in synthetic microniches by selective crosslinking of only cell-laden pre-microgel droplets. Furthermore, we show that we can position cells in the center of the microniches, and that even in protease-sensitive microniches this greatly reduces cell egress. Collectively, we present the development of a versatile protocol that allows for unprecedented efficiency in creation of synthetic protease-sensitive microniches for probing single stem cell fate in 3D.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Técnicas de Cultura de Células / Técnicas Analíticas Microfluídicas / Análise de Célula Única / Microambiente Celular Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Técnicas de Cultura de Células / Técnicas Analíticas Microfluídicas / Análise de Célula Única / Microambiente Celular Tipo de estudo: Diagnostic_studies Limite: Animals Idioma: En Ano de publicação: 2017 Tipo de documento: Article