Title of Oral/Poster Presentation

Comparison of alginate and microcarriers for in vitro modeling of microgravity-induced muscule atrophy

Class

Article

Graduation Year

2017

College

College of Engineering

Department

Biological Engineering Department

Faculty Mentor

Elizabeth Vargis

Presentation Type

Poster Presentation

Abstract

Muscular atrophy, defined as the loss of muscle tissue, is a serious issue for immobilized patients on Earth and in human spaceflight, where microgravity prevents normal muscle loading. Developing countermeasures for atrophy in spaceflight will require extensive screening of promising pharmaceuticals for efficacy, safety, contraindications, and dosage schedule. Due to the cost of spaceflight and limited crew time aboard the International Space Station, high throughput screening of pharmaceuticals in actual microgravity conditions is not economically feasible. We present an optimized ground-based model that induces similar genetic markers of atrophy as seen in spaceflight.

To model the microgravity conditions on the International Space Station, murine C2C12 myoblasts were cultured in a rotary cell culture system. Hydrogel encapsulation was compared against microcarrier beads as a substrate for C2C12 muscle cells. Microcarrier beads are commonly used for suspension culture of adherent cell lines. However, the irregular monolayer that forms on the surface of clustered microcarriers may not accurately model solid muscle tissue. In contrast, encapsulation of differentiated muscle cells in an alginate hydrogel creates a three dimensional tissue model of consistent size and cell density. Changes due to the spaceflight model were characterized qRT-PCR for the atrophy markers MuRF1 and MAFbx. The muscle proteins myosin and tropomyosin were assessed via Western blot.

Encapsulated cells are expected to produce higher levels of MuRF1 and MAFbx than cells cultured on microcarriers. Myosin and tropomyosin levels are also expected to be higher in encapsulated cells, indicating a larger percentage of differentiation. Hydrogel encapsulation increases biosimilarity by using large quantities of pre-differentiated muscle tissue. Simulation of microgravity and spaceflight radiation will provide a valuable platform for drug discovery and an understanding of the progression from normal to disease state.

Location

North Atrium

Start Date

13-4-2017 12:00 PM

End Date

13-4-2017 1:15 PM

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Apr 13th, 12:00 PM Apr 13th, 1:15 PM

Comparison of alginate and microcarriers for in vitro modeling of microgravity-induced muscule atrophy

North Atrium

Muscular atrophy, defined as the loss of muscle tissue, is a serious issue for immobilized patients on Earth and in human spaceflight, where microgravity prevents normal muscle loading. Developing countermeasures for atrophy in spaceflight will require extensive screening of promising pharmaceuticals for efficacy, safety, contraindications, and dosage schedule. Due to the cost of spaceflight and limited crew time aboard the International Space Station, high throughput screening of pharmaceuticals in actual microgravity conditions is not economically feasible. We present an optimized ground-based model that induces similar genetic markers of atrophy as seen in spaceflight.

To model the microgravity conditions on the International Space Station, murine C2C12 myoblasts were cultured in a rotary cell culture system. Hydrogel encapsulation was compared against microcarrier beads as a substrate for C2C12 muscle cells. Microcarrier beads are commonly used for suspension culture of adherent cell lines. However, the irregular monolayer that forms on the surface of clustered microcarriers may not accurately model solid muscle tissue. In contrast, encapsulation of differentiated muscle cells in an alginate hydrogel creates a three dimensional tissue model of consistent size and cell density. Changes due to the spaceflight model were characterized qRT-PCR for the atrophy markers MuRF1 and MAFbx. The muscle proteins myosin and tropomyosin were assessed via Western blot.

Encapsulated cells are expected to produce higher levels of MuRF1 and MAFbx than cells cultured on microcarriers. Myosin and tropomyosin levels are also expected to be higher in encapsulated cells, indicating a larger percentage of differentiation. Hydrogel encapsulation increases biosimilarity by using large quantities of pre-differentiated muscle tissue. Simulation of microgravity and spaceflight radiation will provide a valuable platform for drug discovery and an understanding of the progression from normal to disease state.