Abstract

Current strategies for cartilage cell therapy are mostly based on the use of autologous chondrocytes. However, these cells have limitations of a small number of cells available and of low chondrogenic ability, respectively. Many studies now suggest that fetal stem cells are more plastic than adult stem cells and can therefore more efficiently differentiate into target tissues. This study introduces, efficiency chondrogenic differentiation of fetal cartilage-derived progenitor cells (FCPCs) to adult cells can be achieved using a three-dimensional (3D) spheroid culture method based on silica nanopatterning techniques. In evaluating the issue of silica nano-particle size (Diameter of 300, 750, 1200 nm), each particle size was coated into the well of a 6-well tissue culture plate. FCPCs (2 x 105 cells/well in 6-well plate) were seeded in each well with chondrogenic medium. In this study, the 300 nm substrate that formed multi-spheroids and the 1200 nm substrate that showed spreading were due to the cell-cell adhesion force(via N-cadherin) and cell-substrate(via Integrin) force, the 750 nm substrate that formed the mass-aggregation can be interpreted as the result of cell monolayer formation through cell-substrate force followed by cell-cell contact force contraction. We conclude that our 3D spheroid culture system contributes to an optimization for efficient differentiation of FCPC, offers insight into the mechanism of efficient differentiation of engineered 3D culture system, and has promise for wide applications in regeneration medicine and drug discovery fields.

Highlights

  • The self-repair of articular cartilage (AC) is difficult when it becomes injured due to its low regenerative capacity caused by the lack of blood supply, low cellularity, and a limited number of progenitor cells [1,2,3]

  • The overall homogeneity of substrate coated with silica nano-particles of different size was proved using scanning electron microscope (SEM) (Fig 1)

  • Different results were confirmed from the silica nano-patterned substrate used in this study: mesenchymal stem cells (MSCs) tend to spheroid with larger particle size (350–1200 nm), but it is the multi-spheroids, not one large aggregation (S2 Fig)

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Summary

Introduction

The self-repair of articular cartilage (AC) is difficult when it becomes injured due to its low regenerative capacity caused by the lack of blood supply, low cellularity, and a limited number of progenitor cells [1,2,3]. Autograft cartilage cells have limitations of a small number of cells available and of low chondrogenic ability, respectively. Development of three-dimensional articular cartilage construct using silica nano-patterned substrate shown that stem cells or progenitor cell derived from human fetal tissue is an encouraging cell source for cell therapy and tissue engineering efforts [4, 5]. We have reported that human fetal cartilage progenitor cells (hFCPCs) having high yield, proliferation, multipotent differentiation and maintains the chondrogenic phenotype abilities, in cartilage tissue formation [6]. FCPC can as a novel cell source for cartilage regeneration

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