Rapid isolation, expansion, and differentiation of osteoprogenitors from full-term umbilical cord blood.
There is an urgent clinical requirement for appropriate bone substitutes that can be used for the repair and regeneration of diseased or damaged skeletal tissues. Cell-sourcing limitations in particular have affected progress, largely because of the shortage of accessible tissues capable of yielding sufficient numbers of viable osteoprogenitor cells. Previous work has suggested that umbilical cord blood (UCB) contains circulating progenitor cells (mesenchymal stem cells) capable of osteogenic differentiation, although a comparable number of reports refute this claim. From a screen of more than 20 different culture conditions, we have identified an optimal, simple, and reliable technique to generate, from full-term human UCB, stromal cells with the ability to undergo rapid osteogenic differentiation. By comparing different sorting and culture strategies, we demonstrated that early exposure of mononuclear UCB cells to medium conditioned by osteoblastic cells in the presence of osteogenic supplements and human plasma, markedly increased the frequency of stromal cell growth, the rate of osteogenic differentiation, and their attachment to and spreading on calcium phosphate scaffolds. These findings suggest that full-term UCB may act as an appropriate source of osteoprogenitor cells, which will impact significantly on the development of autologous tissue- engineered bone constructs.
Hutson
Elizabeth L
EL
Biomedical Tissue Research, Department of Biology, University of York, York, United Kingdom. elizabeth.hutson@smith-nephew.com
Boyer
Sam
S
Genever
Paul G
PG
D000135:Acid Phosphatase / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D002130:Calcium Phosphates / Q000378:metabolism / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D002516:Ceramics / Q000378:metabolism / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D006651:Histocytochemistry; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D007797:Laminin / Q000378:metabolism / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D010006:Osteoblasts / Q000378:metabolism; D012516:Osteosarcoma / Q000473:pathology; D016062:Porosity; D011247:Pregnancy; D011263:Pregnancy Trimester, Third; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D000071681:Tartrate-Resistant Acid Phosphatase; D013997:Time Factors
D002130:Calcium Phosphates; D017077:Culture Media, Conditioned; D005353:Fibronectins; D005456:Fluorescent Dyes; D007527:Isoenzymes; D007797:Laminin; D003094:Collagen; C020243:calcium phosphate; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D016650:Fluorescein-5-isothiocyanate