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

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 -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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