Betül SALTIK ÇELEBİ, Beyza YAĞCI GÖKÇINAR

Expansion of human umbilical cord blood hematopoietic progenitors with cord vein pericytes

The vascular niche is a site rich in blood vessels, whereas endothelial cells, pericytes, and smooth muscle cells create a microenvironment that recruits mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs), which is important for stem cell mobilization, proliferation, and differentiation. In this study, CD146 + pericytes were purified and enriched from the human umbilical cord vein. In order to define their direct role in hematopoiesis, we tested the CD146 + pericytes as compared with osteoblasts derived from umbilical cord blood (UCB) MSCs to sustain human UCB hematopoietic progenitor cells in noncontact coculture settings or in culture media previously conditioned (CM) by these cells. The growth of UCB cells was the greatest in pericyte cocultures (2.8-fold vs. the control). The increased growth in pericyte and pericyte CM cultures was largely the result of increased frequency of CD34 + and CD38 + hematopoietic progenitors, CD34 + CD41 + megakaryocyte progenitors, and CD235 + erythroblasts. A total of 29 factors were found to be secreted by pericytes higher than by osteoblasts. The most secreted growth factor by pericytes was vascular endothelial growth factor (1.3-fold). We demonstrate for the first time that human CD146 + perivascular cell coculture and CM are able to directly support the ex vivo maintenance of human hematopoietic progenitor cells.

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Bianco P (2011). Bone and the hematopoietic niche: a tale of two stem cells. Blood 117: 5281-5288.
Birbrair A, Zhang T, Wang ZM, Messi ML, Mintz A, Delbono O (2015). Pericytes at the intersection between tissue regeneration and pathology. Clin Sci (Lond) 128: 81-93.
Bromberg O, Frisch BJ, Weber JM, Porter RL, Civitelli R, Calvi LM (2012). Osteoblastic N-cadherin is not required for microenvironmental support and regulation of hematopoietic stem and progenitor cells. Blood 120: 303-313.
Çelebi B, Mantovani D, Pineault N (2011). Irradiated mesenchymal stem cells improve the ex vivo expansion of hematopoietic progenitors by partly mimicking the bone marrow endosteal environment. J Immunol Methods 370: 93-103.
Çelebi B, Mantovani D, Pineault N (2012). Insulin-like growth factor binding protein-2 and neurotrophin 3 synergize together to promote the expansion of hematopoietic cells ex vivo. Cytokine 58: 327-331.
Corselli M, Chin CJ, Parekh C, Sahaghian A, Wang W, Ge S, Evseenko D, Wang X, Montelatici E, Lazzari L et al. (2013). Perivascular support of human hematopoietic stem/progenitor cells. Blood 121: 2891-2901.
Crisan M, Corselli M, Chen WC, Peault B (2012). Perivascular cells for regenerative medicine. J Cell Mol Med 16: 2851-2860.
Crisan M, Yap S, Casteilla L, Chen CW, Corselli M, Park TS, Andriolo G, Sun B, Zheng B, Zhang L et al. (2008). A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell 3: 301-313.
da Silva CL, Goncalves R, Crapnell KB, Cabral JM, Zanjani ED, Almeida-Porada G (2005). A human stromal-based serum-free culture system supports the ex vivo expansion/maintenance of bone marrow and cord blood hematopoietic stem/progenitor cells. Exp Hematol 33: 828-835.
Darland DC, Massingham LJ, Smith SR, Piek E, Saint-Geniez M, DAmore PA (2003). Pericyte production of cell-associated VEGF is differentiation-dependent and is associated with endothelial survival. Dev Biol 264: 275-288.
Diaz-Flores L, Gutierrez R, Madrid JF, Varela H, Valladares F, Acosta E, Martin-Vasallo P, Diaz-Flores L Jr (2009). Pericytes. Morphofunction, interactions and pathology in a quiescent and activated mesenchymal cell niche. Histol Histopathol 24: 909-969.
Ding L, Saunders TL, Enikolopov G, Morrison SJ (2012). Endothelial and perivascular cells maintain haematopoietic stem cells. Nature 481: 457-462.
Dumont N, Boyer L, Émond H, Çelebi-Saltik B, Pasha R, Bazin R, Mantovani D, Roy DC, Pineault N (2014). Medium conditioned with mesenchymal stromal cell-derived osteoblasts improves the expansion and engraftment properties of cord blood progenitors. Exp Hematol 42: 741-752.e1.
Gökçinar-Yagci B, Özyüncü Ö, Çelebi-Saltik B (2016). Isolation, characterisation and comparative analysis of human umbilical cord vein perivascular cells and cord blood mesenchymal stem cells. Cell Tissue Bank 17: 345-352.
Gökçinar-Yagci B, Uçkan-Çetinkaya D, Çelebi-Saltik B (2015). Pericytes: properties, functions and applications in tissue engineering. Stem Cell Rev 11: 549-559.
Greenbaum AM, Revollo LD, Woloszynek JR, Civitelli R, Link DC (2012). N-cadherin in osteolineage cells is not required for maintenance of hematopoietic stem cells. Blood 120: 295-302.
Hall AP (2006). Review of the pericyte during angiogenesis and its role in cancer and diabetic retinopathy. Toxicol Pathol 34: 763- 775.
He N, Zhang L, Cui J, Li Z (2014). Bone marrow vascular niche: home for hematopoietic stem cells. Bone Marrow Res 2014: 128436.
Himburg HA, Muramoto GG, Daher P, Meadows SK, Russell JL, Doan P, Chi JT, Salter AB, Lento WE, Reya T et al. (2010). Pleiotrophin regulates the expansion and regeneration of hematopoietic stem cells. Nat Med 16: 475-482.
Horwitz ME, Frassoni F (2015). Improving the outcome of umbilical cord blood transplantation through ex vivo expansion or graft manipulation. Cytotherapy 17: 730-738.
Kawano Y, Kobune M, Yamaguchi M, Nakamura K, Ito Y, Sasaki K, Takahashi S, Nakamura T, Chiba H, Sato T et al. (2003). Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells. Blood 101: 532-540.
Kobayashi H, Butler JM, ODonnell R, Kobayashi M, Ding BS, Bonner B, Chiu VK, Nolan DJ, Shido K, Benjamin L et al. (2010). Angiocrine factors from Akt-activated endothelial cells balance self-renewal and differentiation of haematopoietic stem cells. Nat Cell Biol 12: 1046-1056.
Kunisaki Y, Bruns I, Scheiermann C, Ahmed J, Pinho S, Zhang D, Mizoguchi T, Wei Q, Lucas D, Ito K et al. (2013). Arteriolar niches maintain haematopoietic stem cell quiescence. Nature 502: 637-643.
Kunisaki Y, Frenette PS (2014). Influences of vascular niches on hematopoietic stem cell fate. Int J Hematol 99: 699-705.
Levesque JP (2013). A niche in a dish: pericytes support HSC. Blood 121: 2816-2818.
Lilly AJ, Johnson WE, Bunce CM (2011). The haematopoietic stem cell niche: new insights into the mechanisms regulating haematopoietic stem cell behaviour. Stem Cells Int 2011: 274564.
McNiece I, Harrington J, Turney J, Kellner J, Shpall EJ (2004). Ex vivo expansion of cord blood mononuclear cells on mesenchymal stem cells. Cytotherapy 6: 311-317.
Morrison SJ, Scadden DT (2014). The bone marrow niche for haematopoietic stem cells. Nature 505: 327-334.
Potente M, Gerhardt H, Carmeliet P (2011). Basic and therapeutic aspects of angiogenesis. Cell 146: 873-887.
Ribatti D, Nico B, Vacca A (2015). Multiple myeloma as a model for the role of bone marrow niches in the control of angiogenesis. Int Rev Cell Mol Biol 314: 259-282.
Schattner M, Lefebvre P, Mingolelli SS, Goolsby CL, Rademaker A, White JG, Foster D, Green D, Cohen I (1996). Thrombopoietinstimulated ex vivo expansion of human bone marrow megakaryocytes. Stem Cells 14: 207-214.
Sharma MB, Limaye LS, Kale VP (2012). Mimicking the functional hematopoietic stem cell niche in vitro: recapitulation of marrow physiology by hydrogel-based three-dimensional cultures of mesenchymal stromal cells. Haematologica 97: 651-660.
Wang Y, Zhao S (2010). Vascular Biology of the Placenta. San Rafael, CA, USA: Morgan & Claypool Life Sciences.
Winkler EA, Sagare AP, Zlokovic BV (2014). The pericyte: a forgotten cell type with important implications for Alzheimers disease? Brain Pathol 24: 371-386.
Zhu H, Zhang Y, Jin H, Wang Y, Shao X, Kong J, Huang W, Hong Y, Li C, Gao F et al. (2015). Clinical study on 137 cases of unrelated single unit umbilical cord blood hematopoietic stem cell transplantation. Zhonghua Xue Ye Xue Za Zhi 36: 140-143 (in Chinese with English abstract).