Chondrogenesis of human bone marrow-derived mesenchymal stem cells in agarose culture

Anat Rec A Discov Mol Cell Evol Biol. 2004 May;278(1):428-36. doi: 10.1002/ar.a.20010.

Abstract

Mesenchymal stem cells derived from human bone marrow (hBM-MSCs) can differentiate into chondrogenic cells for the potential treatment of injured articular cartilage. To evaluate agarose gels as a supportive material for chondrogenesis of hBM-MSCs, this study examined chondrogenesis of hBM-MSCs in the agarose cultures. Pellet cultures were employed to confirm the chondrogenic potential of the hBM-MSCs that were used in agarose cultures. The hBM-MSCs were seeded in 2% agarose constructs at the initial cell-seeding densities of 3, 6, and 9 x 10(6) cells/ml while each of pellets was formed using 2.5 x 10(5) cells. Chondrogenesis of hBM-MSCs was induced by culturing cell-agarose constructs and pellets for 21 days in the presence of a defined medium containing transforming growth factor beta3 (TGF-beta3). The analysis of reverse transcription-polymerase chain reaction showed that hBM-MSCs of agarose and pellet cultures expressed the chondrogenic markers of collagen type II and aggrecan in the presence of TGF-beta3. The deposition of cartilage-specific macromolecules was detected in both agarose and pellet cultures by histological and immunohistochemical assessments. Chondrogenesis of hBM-MSCs in agarose gels directly correlated with the initial cell-seeding density, with the cell-agarose constructs of higher initial cell-seeding density exhibiting more cartilage-specific gene expressions. This study establishes a basic model for future studies on chondrogenesis of hBM-MSCs using the agarose cultures.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aggrecans
  • Bone Marrow Cells / physiology*
  • Chondrogenesis / physiology*
  • Collagen Type II / metabolism
  • Culture Media
  • DNA Primers
  • Extracellular Matrix Proteins / metabolism
  • Feasibility Studies
  • Histological Techniques
  • Humans
  • Immunohistochemistry
  • Lectins, C-Type
  • Mesenchymal Stem Cells / physiology*
  • Proteoglycans / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sepharose
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta3

Substances

  • Aggrecans
  • Collagen Type II
  • Culture Media
  • DNA Primers
  • Extracellular Matrix Proteins
  • Lectins, C-Type
  • Proteoglycans
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta3
  • Sepharose