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Phenotypic analysis of bovine chondrocytes cultured in 3D collagen sponges: effect of serum substitutes

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Abstract

Repair of damaged cartilage usually requires replacement tissue or substitute material. Tissue engineering is a promising means to produce replacement cartilage from autologous or allogeneic cell sources. Scaffolds provide a three-dimensional (3D) structure that is essential for chondrocyte function and synthesis of cartilage-specific matrix proteins (collagen type II, aggrecan) and sulfated proteoglycans. In this study, we assessed porous, 3D collagen sponges for in vitro engineering of cartilage in both standard and serum-free culture conditions. Bovine articular chondrocytes (bACs) cultured in 3D sponges accumulated and maintained cartilage matrix over 4 weeks, as assessed by quantitative measures of matrix content, synthesis, and gene expression. Chondrogenesis by bACs cultured with Nutridoma as a serum replacement was equivalent or better than control cultures in serum. In contrast, chondrogenesis in insulin-transferrin-selenium (ITS+3) serum replacement cultures was poor, apparently due to decreased cell survival. These data indicate that porous 3D collagen sponges maintain chondrocyte viability, shape, and synthetic activity by providing an environment favorable for high-density chondrogenesis. With quantitative assays for cartilage-specific gene expression and biochemical measures of chondrogenesis in these studies, we conclude that the collagen sponges have potential as a scaffold for cartilage tissue engineering.

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Abbreviations

2D:

two-dimensional

3D:

three-dimensional

AGG:

aggrecan

bACs:

bovine articular chondrocytes

COL I:

type I collagen

COL II:

type II collagen

FBS:

fetal bovine serum

G3PDH:

glyceraldehyde-3-phosphate dehydrogenase

HEPES:

4-(2-hydroxyethyl)piperazine-1-ethansulfonic acid

ITS:

insulin-transferrin-sodium selenite

Nut:

nutridoma

OD:

optical density

RT-PCR:

reverse transcriptase-polymerase chain reaction

s-GAG:

sulfated glycosaminoglycan

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Authors and Affiliations

  1. Department of Orthopedic Surgery, Brigham and Women’s Hospital, Harvard Medical School, 75 Francis Street, 02115, Boston, MA, USA

    Karen E. Yates, Florin Allemann & Julie Glowacki

  2. Skeletal Biology Research Center, Massachusetts General Hospital, Harvard School of Dental Medicine, Boston, MA, USA

    Karen E. Yates, Florin Allemann & Julie Glowacki

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  1. Karen E. Yates
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  2. Florin Allemann
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  3. Julie Glowacki
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Correspondence to Julie Glowacki.

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Yates, K.E., Allemann, F. & Glowacki, J. Phenotypic analysis of bovine chondrocytes cultured in 3D collagen sponges: effect of serum substitutes. Cell Tissue Banking 6, 45–54 (2005). https://doi.org/10.1007/s10561-005-5810-0

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