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Kategorien: Alle - morphology - outcomes - control - results

von Morten Juhl Vor 12 Jahren

268

Jan 2013

The document delves into various aspects of cellular studies, focusing on murine and human nuclei within myotubes. It outlines the methodology employed, including the development of specific cell lines such as C2C12 and ASC-GFP, and the use of cellular stains for observing different cell components.

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Results

Outcomes

Expectations

Stimulated cells

Higher involvement in myotube formation

Alignment, perpendicular to direction of strain

Control

Unlikely >1 human nuclei per myotube

Low degree of fusion (<5%)

Measures

Degree of differentiation

Presence of Myosin

Morphology

No of syncytial nuclei

Human nuclei

Murine nuclei

Size

Directionality

Methodology

Mechanical Stimulation

Uniaxial cyclic tensile strain

Flexcell setup

6 well plates or the new design?

Cell line development

C2C12

C2C12-LifeAct-RFP

C2C12-GFP

ASC-GFP

Cellular stains

Myosin Heavy Chain

Primary [1:500]: Rabbit-anti-myosin

Secondary (10+10 ul/cm2): Zenon Alexa Fluor 647 mouse-anti-rabbit IgG

Cytoplasm

Green Fluorescent Protein

Ex: 487; Em: 509

Human Nuclei

Nuclear envelope

Primary [1:2000]: Monoclonal rabbit-anti-human Lamin

Secondary [1:400]: Alexa Fluor 555 conjugated goat-anti-rabbit

All Nuclei

DNA

Hoechst 33342

Jan 2013

The document delves into various aspects of cellular studies, focusing on murine and human nuclei within myotubes. It outlines the methodology employed, including the development of specific cell lines such as C2C12 and ASC-GFP, and the use of cellular stains for observing different cell components.