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Abstract

OBJECTIVE: To analyze the growth patterns and differentiation of human septal chondrocyte monolayers of different seeding densities.

STUDY DESIGN: Chondrocytes from 8 donors were plated at densities ranging from 20,000 cells/cm² (high density) to 300 cells/cm² (very low density). Confluency, cellularity, and glycosaminoglycan content were determined from days 1 to 15.

RESULTS: Confluency was attained at 5.8, 8.3, 11.0, and 14.8 days for high-, intermediate-, low-, and very low-density monolayers, respectively (P >0.001). Regression growth curves showed typical lag, logarithmic, and stationary phases. Confluent monolayers attained similar cellularity (power = 0.94) and differentiation (power = 0.88), regardless of initial density.

CONCLUSIONS: Human septal chondrocyte monolayers reach confluency from very low initial densities. Growth patterns, cellularity, and differentiation are similar to other starting densities.

SIGNIFICANCE: Very low-density monolayers expanded cell number 838-fold in 1 passage and therefore are sufficient for tissue-engineering purposes. This is important because of the requirement of maintaining differentiation and the limitation of small tissue harvest specimens.

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References

Vacanti

Upton

Tissue engineered morphogenesis of cartilage and bone by means of cell transplantation using synthetic biodegradable polymer matrices. Clin Plast Surg 1994; 21: 445–62.

2 Brittberg

Lindahl

Nilsson

, Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med 1994; 331: 889–95.

Benya

Shaffer

Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 1982; 30: 215–24.

4 Vacanti

Langer

Schloo

, Synthetic Polymers seeded with chondrocytes provide a template for new cartilage formation. Plast Reconstr Surg 1991; 88: 753–9.

5 Lee

Moon

Choi

, Tissue engineered tracheal prosthesis with acceleratedly cultured homologous chondrocytes as an alternative of tracheal reconstruction. J Cardiovasc Surg 2002; 43: 275–9.

van Osch

van der Veen

Verwoerd-Verhoef

HL.

In vitro redifferentiation of culture-expanded rabbit and human auricular chondrocytes for cartilage reconstruction. Plast Reconstr Surg 2001; 107: 433–40.

7 Rotter

Tobias

Lebl

, Age-related changes in the composition and mechanical properties of human nasal cartilage. Arch Biochem Biophys 2002; 403: 132–40.

Koch

Gorti

GK.

Tissue engineering with chondrocytes. Facial Plast Surg 2002; 18: 59–68.

9 Pacifici

Boettiger

Roby

, Transformation of chondroblasts by Rous sarcoma virus and synthesis of the sulfated proteoglycan matrix. Cell 1977; 11: 891–9.

10.

10 von der Mark

Verena

von der Mark

, Relationship between cell shape and type of collagen synthesized as chondrocytes lose their cartilage phenotype in culture. Nature 1977; 267: 531–2.

11.

11 Homicz

Schumacher

Sah

, Effects of serial expansion of septal chondrocytes on tissue-engineered neocartilage composition. Otolaryngol Head Neck Surg 2002; 127: 398–408.

12.

12 Kim

Sah

Doong

, Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem 1988; 174: 168–76.

13.

13 Puelacher

Kim

Vacanti

, Tissue-engineered growth of cartilage: the effects of varying the concentration of chondrocytes seeded onto synthetic polymer matrices. Int J Oral Maxillofac Surg 1994; 23: 49–53.

14.

14 Marijnissen

van Osch

Aigner

, Tissue-engineered cartilage using serially passaged articular chondrocytes. Chondrocytes in alginate, combined in vivo with a synthetic (E210) or biologic biodegradable carrier (DBM). Biomaterials 2000; 21: 571–80.

15.

15 Kuettner

Pauli

Gall

, Synthesis of cartilage matrix by mammalian chondrocytes in vitro. I. solation, culture characteristics, and morphology. J Cell Biol 1982; 93: 743–50.

16.

Watt

FM.

Effect of seeding density on stability of the differentiated phenotype of pig articular chondrocytes in culture. J Cell Sci 1988; 89: 373–8.

17.

17 Yaeger

Masi

de Ortiz

JLB

, Synergistic action of transforming growth factor-β and insulin-like growth factor-I induces expression of type II collagen and aggrecan genes in adult human articular chondrocytes. Exp Cell Res 1997; 237: 318–25.

18.

18 Trippel

Wroblewski

Makower

, Regulation of growthplate chondrocytes by insulin-like growth factor-I and basic fibroblast growth factor. J Bone Joint Surg Am 1994; 75: 177–89.

19.

19 Kuriwaka

Ochi

Uchio

, Optimum combination of monolayer and three-dimensional cultures for cartilage-like tissue engineering. Tissue Eng 2003; 9: 41–9.

20.

Tsoularis

Wallace

Analysis of logistic growth models. Math Biosci 2002; 179: 21–55.

Growth and Phenotype of Low-Density Nasal Septal Chondrocyte Monolayers

Abstract

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