title: Gene expression changes in an in vitro model of chondroinduction: a comparison of two methods.
abstract: There are many useful technologies to describe patterns of gene expression that occur during tissue repair and regeneration. Results from different methods used in one experimental setting are not often compared. In this case study of chondrogenesis, we compare two methods to identify differentially expressed genes, representational difference analysis and targeted macroarray analysis, as a model for investigating genes that may be relevant to tissue repair. We sought to identify genes whose expression was altered when human dermal fibroblasts were cultured in a three-dimensional, porous collagen sponge with the chondroinductive agent, demineralized bone. Both representational difference analysis and macroarray experiments revealed several functional families of genes as up-regulated or down-regulated in chondroinduced fibroblasts. An advantage of representational difference analysis is that altered expression of specific mRNA transcripts can be revealed. In this example, representational difference analysis uncovered the up-regulation of a specific transcript of Wnt5a in fibroblasts cultured with demineralized bone. Representational difference analysis is limited, however, as there can be false negatives for genes not readily amplified by polymerase chain reaction. We conclude that small arrays containing functional classes of genes can be used to ask specific, hypothesis-driven questions at minimal cost. It may be prudent, however, to use more than one method to survey differences in gene expression in order to validate and expand findings.
authors.affiliation: Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, 75 Francis Street, Boston, MA 02115, USA. kyates@partners.org
authors.forename: Karen E
authors.initials: KE
authors.lastname: Yates
authors.forename: Julie
authors.initials: J
authors.lastname: Glowacki
mesh_terms: D001842:Bone and Bones / Q000502:physiology*; D018929:Cell Culture Techniques; D020219:Chondrogenesis / Q000235:genetics*; D003094:Collagen / Q000502:physiology*; D020405:Dermis / Q000166:cytology / Q000502:physiology; D005347:Fibroblasts / Q000502:physiology*; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011208:Powders; D015398:Signal Transduction / Q000235:genetics
chemical_list: D011208:Powders; D003094:Collagen