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Stem Cells 2005;23:220-229 www.StemCells.com
© 2005 AlphaMed Press

Human Umbilical Cord Perivascular (HUCPV) Cells: A Source of Mesenchymal Progenitors

Rahul Sarugaser, David Lickorish, Dolores Baksh, M. Morris Hosseini, John E. Davies

Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada

Key Words. Mesenchymal progenitors • Umbilical cord • Allogeneic cells • Major histocompatibility complexes • Cryopreservation • Therapeutic dose

Correspondence: J. E. Davies, B.D.S., D.Sc., Institute of Biomaterials and Biomedical Engineering, University of Toronto, 4 Taddle Creek Road, Room 407, Toronto, ON M5S 3G9, Canada. Telephone: 416-978-1471; Fax: 416-946-5639 ; e-mail: davies{at}ecf.utoronto.ca; Website: http://www.ecf.utoronto.ca/~bonehead

We describe the isolation of a nonhematopoietic (CD45–, CD34–, SH2+, SH3+, Thy-1+, CD44+) human umbilical cord perivascular (HUCPV) cell population. Each HUCPV cell harvest (2–5 x 106, depending on the length of cord available) gave rise to a morphologically homogeneous fibroblastic cell population, which expressed {alpha}-actin, desmin, vimentin, and 3G5 (a pericyte marker) in culture. We determined the colony-forming unit-fibro-blast (CFU-F) frequency of primary HUCPV cells to be 1:333 and the doubling time, which was 60 hours at passage 0 (P0), decreased to 20 hours at P2. This resulted in a significant cell expansion, producing over 1010 HUCPV cells within 30 days of culture. Furthermore, HUCPV cells cultured in nonosteogenic conditions contained a subpopulation that exhibited a functional osteogenic phenotype and elaborated bone nodules. The frequency of this CFU-osteogenic subpopulation at P1 was 2.6/105 CFU-F, which increased to 7.5/105 CFU-F at P2. Addition of osteogenic supplements to the culture medium resulted in these frequencies increasing to 1.2/104 and 1.3/104 CFU-F, respectively, for P1 and P2. CFU-O were not seen at P0 in either osteogenic or non-osteogenic culture conditions, but P0 HUCPV cells did contain a 20% subpopulation that presented neither class I nor class II cell-surface major histocompatibility complexes (MHC–/–). This population increased to 95% following passage and cryopreservation (P5). We conclude that, due to their rapid doubling time, high frequencies of CFU-F and CFU-O, and high MHC–/– phenotype, HUCPV cells represent a significant source of cells for allogeneic mesenchymal cell-based therapies.




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