title: Development of methods for studying the differentiation of human mesenchymal stem cells under cyclic compressive strain.
abstract: Human mesenchymal stem cells (hMSC) have numerous potential advantages over terminally differentiated cells and embryonic stem cells for use in tissue engineering applications. The aims of this study were to develop methods to test the hypothesis that hMSC could be differentiated using cyclic compressive strain alone. hMSC were successfully isolated, purified using D7-FIB antibody, cloned, and characterized. The cells were subsequently analyzed using fluorescence-activated cell sorting using a panel of antibodies and differentiation into multiple cell lineages. D7FIB-positive cells were then seeded into collagen-alginate scaffolds and subjected to 10% or 15% cyclic compressive strain for 4 out of 24 hours for up to 21 days in a bespoke servo-assisted displacement-controlled device. Cells were analyzed using adenosine triphosphate assay to determine cell number, live-dead cell assay, and quantitative real-time polymerase chain reaction at 7 and 21 days. Cloned D7-FIB-positive hMSCs showed evidence of differentiation to an osteogenic lineage under 10% cyclic compressive strain alone (core binding factor alpha 1 (CBFA-1) was significantly upregulated at 7 and 21 days by a factor of 18.3 and 32.2, respectively) and to an osteo-chondrogenic lineage under 15% cyclic compressive strain alone (increased expression of CBFA-1, Sox9, and aggrecan). A combination of a composite viscoelastic scaffold and controlled cyclic compressive strain may be useful for study of the differentiation of MSC.
authors: Faculty of Biological Sciences, Institute of Medical & Biological Engineering, University of Leeds, Leeds, United Kingdom.
authors: Efstathios
authors: E
authors: Michalopoulos
authors: Richard L
authors: RL
authors: Knight
authors: Sotirios
authors: S
authors: Korossis
authors: John N
authors: JN
authors: Kearney
authors: John
authors: J
authors: Fisher
authors: Eileen
authors: E
authors: Ingham
mesh_terms: D000368:Aged; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000464:Alginates / Q000494:pharmacology; D000818:Animals; D001696:Biomechanical Phenomena / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008875:Middle Aged; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D013314:Stress, Mechanical; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D053674:Aggrecans; D000464:Alginates; D006603:Hexuronic Acids; D047495:PPAR gamma; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D020723:Glucuronic Acid; D003094:Collagen