Spheroid Coculture of Hematopoietic Stem/Progenitor Cells and Monolayer Expanded Mesenchymal Stem/Stromal Cells in Polydimethylsiloxane Microwells Modestly Improves In Vitro Hematopoietic Stem/Progenitor Cell Expansion

被引:44
作者
Futrega, Kathryn [1 ]
Atkinson, Kerry [1 ]
Lott, William B. [1 ]
Doran, Michael R. [1 ,2 ]
机构
[1] Queensland Univ Technol, Translat Res Inst, Stem Cell Therapies Lab, 37 Kent St, Brisbane, Qld 4102, Australia
[2] Univ Queensland, Mater Res Inst, Translat Res Inst, Brisbane, Qld, Australia
基金
英国医学研究理事会;
关键词
co-culture; cord blood hematopoietic stem cells; engraftment; mesenchymal stem cells; microwell; spheroid culture; EX-VIVO EXPANSION; HUMAN CORD BLOOD; STEM-CELLS; PROGENITOR CELLS; RETINOIC ACID; STROMAL CELLS; ENDOTHELIAL-CELLS; GENE-EXPRESSION; BONE; CULTURES;
D O I
10.1089/ten.tec.2016.0329
中图分类号
Q813 [细胞工程];
学科分类号
摘要
While two-dimensional (2D) monolayers of mesenchymal stem/stromal cells (MSCs) have been shown to enhance hematopoietic stem/progenitor cell (HSPC) expansion in vitro, expanded cells do not engraft long term in human recipients. This outcome is attributed to the failure of 2D culture to recapitulate the bone marrow (BM) niche signal milieu. Herein, we evaluated the capacity of a novel three-dimensional (3D) coculture system to support HSPC expansion in vitro. A high-throughput polydimethylsiloxane (PDMS) microwell platform was used to manufacture thousands of uniform 3D multicellular coculture spheroids. Relative gene expression in 3D spheroid versus 2D adherent BM-derived MSC cultures was characterized and compared with literature reports. We evaluated coculture spheroids, each containing 25-400 MSCs and 10 umbilical cord blood (CB)-derived CD34(+) progenitor cells. At low exogenous cytokine concentrations, 2D and 3D MSC coculture modestly improved overall hematopoietic cell and CD34(+) cell expansion outcomes. By contrast, a substantial increase in CD34(+) CD38(-) cell yield was observed in PDMS microwell cultures, regardless of the presence or absence of MSCs. This outcome indicated that CD34(+) CD38(-) cell culture yield could be increased using the microwell platform alone, even without MSC coculture support. We found that the increase in CD34(+) CD38(-) cell yield observed in PDMS microwell cultures did not translate to enhanced engraftment in NOD/SCID gamma (NSG) mice or a modification in the relative human hematopoietic lineages established in engrafted mice. In summary, there was no statistical difference in CD34(+) cell yield from 2D or 3D cocultures, and MSC coculture support provided only modest benefit in either geometry. While the high-throughput 3Dmicrowell platform may provide a useful model system for studying cells in coculture, further optimization will be required to generate HSPC yields suitable for use in clinical applications.
引用
收藏
页码:200 / 218
页数:19
相关论文
共 79 条
  • [11] Minireview: The Stem Cell Next Door: Skeletal and Hematopoietic Stem Cell "Niches" in Bone
    Bianco, Paolo
    [J]. ENDOCRINOLOGY, 2011, 152 (08) : 2957 - 2962
  • [12] Aryl Hydrocarbon Receptor Antagonists Promote the Expansion of Human Hematopoietic Stem Cells
    Boitano, Anthony E.
    Wang, Jian
    Romeo, Russell
    Bouchez, Laure C.
    Parker, Albert E.
    Sutton, Sue E.
    Walker, John R.
    Flaveny, Colin A.
    Perdew, Gary H.
    Denison, Michael S.
    Schultz, Peter G.
    Cooke, Michael P.
    [J]. SCIENCE, 2010, 329 (5997) : 1345 - 1348
  • [13] Cook MM, 2012, TISSUE ENG PART C-ME, V18, P319, DOI [10.1089/ten.TEC.2011.0159, 10.1089/ten.tec.2011.0159]
  • [14] Perivascular support of human hematopoietic stem/progenitor cells
    Corselli, Mirko
    Chin, Chee Jia
    Parekh, Chintan
    Sahaghian, Arineh
    Wang, Wenyuan
    Ge, Shundi
    Evseenko, Denis
    Wang, Xiaoyan
    Montelatici, Elisa
    Lazzari, Lorenza
    Crooks, Gay M.
    Peault, Bruno
    [J]. BLOOD, 2013, 121 (15) : 2891 - 2901
  • [15] BMP signalling differentially regulates distinct haematopoietic stem cell types
    Crisan, Mihaela
    Kartalaei, Parham Solaimani
    Vink, Chris
    Yamada-Inagawa, Tomoko
    Bollerot, Karine
    van IJcken, Wilfred
    van der Linden, Reinier
    Lopes, Susana M. Chuva de Sousa
    Monteiro, Rui
    Mummery, Christine
    Dzierzak, Elaine
    [J]. NATURE COMMUNICATIONS, 2015, 6
  • [16] Peptide agonist of the thrombopoietin receptor as potent as the natural cytokine
    Cwirla, SE
    Balasubramanian, P
    Duffin, DJ
    Wagstrom, CR
    Gates, CM
    Singer, SC
    Davis, AM
    Tansik, RL
    Mattheakis, LC
    Boytos, CM
    Schatz, PJ
    Baccanari, DP
    Wrighton, NC
    Barrett, RW
    Dower, WJ
    [J]. SCIENCE, 1997, 276 (5319) : 1696 - 1699
  • [17] Ex vivo expansion of human hematopoietic stem and progenitor cells
    Dahlberg, Ann
    Delaney, Colleen
    Bernstein, Irwin D.
    [J]. BLOOD, 2011, 117 (23) : 6083 - 6090
  • [18] Osteoblasts and Bone Marrow Mesenchymal Stromal Cells Control Hematopoietic Stem Cell Migration and Proliferation in 3D In Vitro Model
    de Barros, Ana Paula D. N.
    Takiya, Christina M.
    Garzoni, Luciana R.
    Leal-Ferreira, Mona Lisa
    Dutra, Helio S.
    Chiarini, Luciana B.
    Meirelles, Maria Nazareth
    Borojevic, Radovan
    Rossi, Maria Isabel D.
    [J]. PLOS ONE, 2010, 5 (02):
  • [19] Cord-Blood Engraftment with Ex Vivo Mesenchymal-Cell Coculture
    de Lima, Marcos
    McNiece, Ian
    Robinson, Simon N.
    Munsell, Mark
    Eapen, Mary
    Horowitz, Mary
    Alousi, Amin
    Saliba, Rima
    McMannis, John D.
    Kaur, Indreshpal
    Kebriaei, Partow
    Parmar, Simrit
    Popat, Uday
    Hosing, Chitra
    Champlin, Richard
    Bollard, Catherine
    Molldrem, Jeffrey J.
    Jones, Roy B.
    Nieto, Yago
    Andersson, Borje S.
    Shah, Nina
    Oran, Betul
    Cooper, Laurence J. N.
    Worth, Laura
    Qazilbash, Muzaffar H.
    Korbling, Martin
    Rondon, Gabriela
    Ciurea, Stefan
    Bosque, Doyle
    Maewal, Ila
    Simmons, Paul J.
    Shpall, Elizabeth J.
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 2012, 367 (24) : 2305 - 2315
  • [20] Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution
    Delaney, Colleen
    Heimfeld, Shelly
    Brashem-Stein, Carolyn
    Voorhies, Howard
    Manger, Ronald L.
    Bernstein, Irwin D.
    [J]. NATURE MEDICINE, 2010, 16 (02) : 232 - U143