Coculture of meniscus cells and mesenchymal stem cells in simulated microgravity

William M Weiss,Adetola B Adesida,Aillette Mulet-Sierra,Melanie Kunze,Nadr M Jomha

doi:10.1038/s41526-017-0032-x

Abstract

Simulated microgravity has been shown to enhance cartilaginous matrix formation by chondrocytes and chondrogenesis of mesenchymal stem cells (MSCs). Similarly, coculture of primary chondrocytes with MSCs has been shown as a strategy to simultaneously retain the differentiated phenotype of chondrocytes and enhance cartilaginous matrix formation. In this study, we investigated the effect of simulated microgravity on cocultures of primary human meniscus cells and adipose-derived MSCs. We used biochemical, qPCR, and immunofluorescence assays to conduct our investigation. Simulated microgravity significantly enhanced cartilaginous matrix formation in cocultures of primary meniscus cells and adipose-derived MSCs. The enhancement was accompanied by increased hypertrophic differentiation markers, COL10A1 and MMP-13, and suppression of hypertrophic differentiation inhibitor, gremlin 1 (GREM1).

Highlights

The menisci of the knee are a pair of fibrocartilaginous tissues.[1]They primarily serve as mechanical load distributors within the knee joint.[2]
We determined the clonogenicity of the mononucleated cells (MNCs) to Colony forming characteristics and immuno-phenotype of adipose stem cells (ASC)
We explored the possibility that cell proliferation may Chondro-induction is enhanced in simulated microgravity (SMG) cocultures of primary meniscus cells (MC) and ASC

Summary

Introduction

The menisci of the knee are a pair of fibrocartilaginous tissues.[1] They primarily serve as mechanical load distributors within the knee joint.[2] Traumatic tears in the avascular region of the tissue are common and do not heal.[1] Treatment options for these tears are currently limited to partial meniscectomy.[3,4] partial meniscectomy is a major risk factor for the early development of knee osteoarthritis.[3,4] Tissue engineering using cells presents a potential option to create functional tissues to replace damaged meniscus.[5,6,7,8,9,10,11,12,13,14,15] Primary meniscus cells (MC) are the ideal cell sources as they closely resemble the in vivo phenotype of MC and can form the biomechanically functional extracellular matrix (ECM) of the meniscus.[1,16,17] Obtaining sufficient numbers of primary MC for meniscus tissue engineering is often impossible and impractical.[18,19] To circumvent this limitation, primary MC are expanded in in vitro culture. Hypoxia stimulated chondrogenic differentiation of bone marrow-derived MSCs by induction of TGF-β1 gene expression and protein production.[33]

Objectives

Methods

Results

Conclusion