Specification of chondrocytes and cartilage tissues from embryonic stem cells

TY - JOUR

T1 - Specification of chondrocytes and cartilage tissues from embryonic stem cells

AU - Craft, April M.

AU - Ahmed, Nazish

AU - Rockel, Jason S.

AU - Baht, Gurpreet S.

AU - Alman, Benjamin A.

AU - Kandel, Rita A.

AU - Grigoriadis, Agamemnon E.

AU - Keller, Gordon M.

PY - 2013/6/15

Y1 - 2013/6/15

N2 - Osteoarthritis primarily affects the articular cartilage of synovial joints. Cell and/or cartilage replacement is a promising therapy, provided there is access to appropriate tissue and sufficient numbers of articular chondrocytes. Embryonic stem cells (ESCs) represent a potentially unlimited source of chondrocytes and tissues as they can generate a broad spectrum of cell types under appropriate conditions in vitro. Here, we demonstrate that mouse ESC-derived chondrogenic mesoderm arises from a Flk-1(-)/Pdgfr alpha(+) (F-P+) population that emerges in a defined temporal pattern following the development of an early cardiogenic F-P+ population. Specification of the late-arising F-P+ population with BMP4 generated a highly enriched population of chondrocytes expressing genes associated with growth plate hypertrophic chondrocytes. By contrast, specification with Gdf5, together with inhibition of hedgehog and BMP signaling pathways, generated a population of non-hypertrophic chondrocytes that displayed properties of articular chondrocytes. The two chondrocyte populations retained their hypertrophic and non-hypertrophic properties when induced to generate spatially organized proteoglycan-rich cartilage-like tissue in vitro. Transplantation of either type of chondrocyte, or tissue generated from them, into immunodeficient recipients resulted in the development of cartilage tissue and bone within an 8-week period. Significant ossification was not observed when the tissue was transplanted into osteoblast-depleted mice or into diffusion chambers that prevent vascularization. Thus, through stage-specific manipulation of appropriate signaling pathways it is possible to efficiently and reproducibly derive hypertrophic and non-hypertrophic chondrocyte populations from mouse ESCs that are able to generate distinct cartilage-like tissue in vitro and maintain a cartilage tissue phenotype within an avascular and/or osteoblast-free niche in vivo.

AB - Osteoarthritis primarily affects the articular cartilage of synovial joints. Cell and/or cartilage replacement is a promising therapy, provided there is access to appropriate tissue and sufficient numbers of articular chondrocytes. Embryonic stem cells (ESCs) represent a potentially unlimited source of chondrocytes and tissues as they can generate a broad spectrum of cell types under appropriate conditions in vitro. Here, we demonstrate that mouse ESC-derived chondrogenic mesoderm arises from a Flk-1(-)/Pdgfr alpha(+) (F-P+) population that emerges in a defined temporal pattern following the development of an early cardiogenic F-P+ population. Specification of the late-arising F-P+ population with BMP4 generated a highly enriched population of chondrocytes expressing genes associated with growth plate hypertrophic chondrocytes. By contrast, specification with Gdf5, together with inhibition of hedgehog and BMP signaling pathways, generated a population of non-hypertrophic chondrocytes that displayed properties of articular chondrocytes. The two chondrocyte populations retained their hypertrophic and non-hypertrophic properties when induced to generate spatially organized proteoglycan-rich cartilage-like tissue in vitro. Transplantation of either type of chondrocyte, or tissue generated from them, into immunodeficient recipients resulted in the development of cartilage tissue and bone within an 8-week period. Significant ossification was not observed when the tissue was transplanted into osteoblast-depleted mice or into diffusion chambers that prevent vascularization. Thus, through stage-specific manipulation of appropriate signaling pathways it is possible to efficiently and reproducibly derive hypertrophic and non-hypertrophic chondrocyte populations from mouse ESCs that are able to generate distinct cartilage-like tissue in vitro and maintain a cartilage tissue phenotype within an avascular and/or osteoblast-free niche in vivo.

KW - Cartilage

KW - Chondrocyte

KW - Embryonic stem cell

KW - Induced pluripotent stem cell

KW - Paraxial

KW - Somite

KW - ARTICULAR-CARTILAGE

KW - IN-VITRO

KW - MOUSE EMBRYO

KW - CHONDROGENIC DIFFERENTIATION

KW - SKELETAL DEVELOPMENT

KW - PRIMITIVE STREAK

KW - PROGENITOR CELLS

KW - SOMITE FORMATION

KW - SYNOVIAL JOINT

KW - EXPRESSION

U2 - 10.1242/dev.087890

DO - 10.1242/dev.087890

M3 - Article

SN - 0950-1991

VL - 140

SP - 2597

EP - 2610

JO - Development

JF - Development

IS - 12

ER -