Hydrostatic pressure promotes chondrogenic differentiation and microvesicle release from human embryonic and bone marrow stem cells.

Lu Luo,Alicia J El Haj,David A Hoey,Sophie C Cox,Kenny L Man,Mathieu Brunet,Nicola C Foster,Susan J Kimber

doi:10.1002/biot.202100401

Abstract

Mechanical stimulation plays in an important role in regulating stem cell differentiation and their release of extracellular vesicles (EVs). In this study, effects of low magnitude hydrostatic pressure (HP) on the chondrogenic differentiation and microvesicle release from human embryonic stem cells (hESCs) and human bone marrow stem cells (hBMSCs) are examined. hESCs were differentiated into chondroprogenitors and then embedded in fibrin gels and subjected to HP (270kPa, 1Hz, 5days per week). hBMSC pellets were differentiated in chondrogenic media and subjected to the same regime. HP significantly enhanced ACAN expression in hESCs. It also led to a significant increase in DNA content, sGAG content and total sGAG/DNA level in hBMSCs. Furthermore, HP significantly increased microvesicle protein content released from both cell types. These results highlight the benefit of HP bioreactor in promoting chondrogenesis and EV production for cartilage tissue engineering.

Highlights

Osteoarthritis (OA), a disease characterized by the degeneration and loss of articular cartilage, affects at least 250 million people globally [1]
We examined effects of low magnitude hydrostatic pressure (HP) on the chondrogenic differentiation and microvesicle release from human embryonic stem cells and human bone marrow stem cells. hESCs were differentiated into chondroprogenitors and embedded in fibrin gels and subjected to HP (270 kPa, 1Hz, 5 days/week). hBMSC pellets were differentiated in chondrogenic media and subjected to the same regime
After 14 days of directed differentiation, the hESC marker OCT4 and NANOG significantly reduced whereas the chondrogenic markers including Sox9, COL2A1 and ACAN all increased significantly (Supplementary Fig.1A)

Summary

Introduction

Osteoarthritis (OA), a disease characterized by the degeneration and loss of articular cartilage, affects at least 250 million people globally [1]. The procurement of stem cells from the bone marrow is a surgical procedure associated with pain and risk of complications. There are thousands of human embryonic stem cells (ESCs) lines available from surplus embryos donated from in vitro fertilization procedures and a number of these are of clinical grade [12]. They are pluripotent and possess unlimited selfrenewal capacity, may form an alternative allogeneic cell source for cartilage tissue engineering applications. The behaviour of ESCs has been investigated in a number of naturally derived hydrogels, such as agarose [13], hyaluronic acid [14, 15] and fibrin gels [16, 17], for cartilage tissue engineering applications

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