Chondrogenic Differentiation of Human Adipo-Derived Precursor Cells.

Jeong, Jae Ho

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Original Article

Journal of the Korean Society of Plastic and Reconstructive Surgeons 2000;27(2):136-142.
Published online March 1, 2000.

Chondrogenic Differentiation of Human Adipo-Derived Precursor Cells.

Jae Ho Jeong

Department of Plastic & Reconstructive Surgery, Yeungnam University Medical Center, Taegu, Korea.

Abstract

According to recent advances in tissue engineering, several tissue types of mesodermal origin are successfully engineered In order to engineer connective tissues, a clinically practical and readily available source of cells must be identified Based on recent work relating to the existence of undifferentiated multipotential stem cells, they exist not only in bone marrow but also in many other connective tissue stromal components. We wondered if human fat tissue might also contain cells with developmental plasticity. More specifically, the purpose of this study was to induce cells isolated from the stromal fraction of human adipose tissue along the chondrogenic lineage in-vitro. Human adiopse tissue was harvested by conventional liposuction procedure. Collagenase type I was used for adipose tissue dissociation and stromal cell suspension was collected The cells were basically cultured in Opti-mem"". For chondrogenic digerentiation, a special culture condition, named micromass culture technique, was utilized and the media was supplemented with fetal bovine serum, L-ascorbic acid, ethanol, TGF-g and antibioticg antimycotic agents. The cells were cultured at 37C incubator with 5% COz for 3 weeks. For detection of chondroblast and/or cartilage matrix, alcian blue (pH 1.0) staining, alkaline phosphatase staining, indirect immunofluorescent labeling using ExtrAvidin'-FIZC conjugate were done. Scanning and transmission electron microscopic images were also taken for ultramicroscopic evaluation of the cells. Blue-colored positive reaction was detected in alcian blue staining, which means presence of sulfated mucin ('cartilage matrix). The cells expressed strong reaction of the tissue nonspecific isoform of the enzyme alkaline phosphatase. Bright fluorescence was detected in indirect immunofluorescent labeling representing the presence of collagen type II in the matrix, which is a specific component of cartilage. Based on our results, it can be concluded that human adipose tissue contains certain population of undigerentiated precursor cells, which could be differentiated into several connective tissue cell lineages. This population of cells may be used as an important future source for cellular building blocks for tissue engineering.

Keywords: Chondrogenesis; Tissue engineering; Adipcse tissue

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