Examining the Effects of Passaging on Murine Tenocytes 

Researcher(s)

  • Christian Le, Biomedical Engineering, University of Delaware

Faculty Mentor(s)

  • Justin Parreno, Biological Sciences, University of Delaware

Abstract

 

Authors: Christian Le, Rylee King, Justin Parreno

Tendon is a fibrous connective tissue composed of extracellular matrix proteins (predominantly collagen). Tenocytes, the main resident cells of tendons, maintain tendon homeostasis. Tendon injuries often heal with scar tissue as a result of tendon fibrosis. This pathological process is driven by the activation and persistence of myofibroblasts. While myofibroblasts play a crucial role in matrix remodeling through deposition and contraction, their unremitting activation leads to excessive scar formation and tissue thickening. The extracellular matrix (ECM) scar tissue formed by myofibroblasts do not have the same biomechanical properties as native tendon, leading to the loss of mechanical strength and increasing the frequency of re-injury. Therefore, advancing tendon tissue engineering is crucial for creating treatments to restore native tendon functions.

 

One concern for engineering tendon tissue is acquiring sufficient cell numbers through growth. Expansion of isolated tenocytes has potential; however, the in vitro proliferative capacity of tenocytes is not fully clear. The stability of the tenocyte phenotype following culture expansion is also not completely known. Therefore, it is crucial to characterize the monolayer expansion potential of tenocytes. In this study, we investigate the phenotype of native and passaged tenocytes, determining whether passaged tenocytes could maintain their matrix molecule expression for tissue engineering purposes. 

 

Based on previous literature, we hypothesize that tenocytes will maintain a tenogenic phenotype with passaging. Using RT-PCR, we will determine whether the passaged tenocytes maintain tenogenic markers, including fibronectin, alpha-smooth muscle actin, and collagen type I. Comparing expression between passages and native tendon will allow us to determine if the tenocytes maintain their phenotype over multiple passages and provide information on how cultured cells differ from native tendons. Understanding the phenotypic stability of tenocytes will allow for determination of the potential of using passaged tenocytes for tissue engineering purposes.