Effect of heparin addition on expansion of cord blood hematopoietic progenitor cells in three-dimensional coculture with stromal cells in nonwoven fabrics.
Primary human cord blood mononuclear cells (CB MNCs) were inoculated into layers of primary human bone marrow stromal cells prepared in a nonwoven fabric porous carrier [three dimensional (3-D)] or on a dish [two dimensional (2-D)] using a cytokine-free medium and were cultured for 7 days with or without the addition of heparin. The number of progenitor cells increased threefold during the 3-D coculture, whereas it decreased in the 2-D culture. Heparin addition to the 3-D coculture further increased the number of progenitors twofold, whereas the addition of desulfated heparin had no effect. The heparin effect was also observed in a 3-D culture of CB MNCs without stromal cells when conditioned medium was employed. The coating of the carrier with N-(O-beta-(6-O-sulfogalactopyranosyl)-6-oxyhexyl)-3,5-bis (dodecyloxy)-benzamide instead of heparin addition also increased the number of progenitor cells in the 3-D culture of CB MNCs without stromal cells when the conditioned medium was employed. The 3-D coculture constructed with nonwoven fabrics and stromal cells was clearly superior to the 2-D culture because of the expansion of CB hematopoietic progenitor cells without cytokine addition. Heparin addition to the 3-D coculture further increased the number of progenitor cells, which may result from a synergistic effect of soluble cytokines produced by stromal cells with the sulfur group of heparin.
Okamoto
Toru
T
International Center for Biotechnology, Osaka University, Osaka, Japan.
Takagi
Mutsumi
M
Soma
Toshihiro
T
Ogawa
Hiroyasu
H
Kawakami
Manabu
M
Mukubo
Masaaki
M
Kubo
Kazusuke
K
Sato
Reiko
R
Toma
Kazunori
K
Yoshida
Toshiomi
T
D000818:Animals; D001672:Biocompatible Materials; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002478:Cells, Cultured; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D017077:Culture Media, Conditioned; D005312:Fetal Blood / Q000166:cytology; D006410:Hematopoiesis / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D011336:Probability; D012680:Sensitivity and Specificity; D023822:Tissue Engineering
D001672:Biocompatible Materials; D017077:Culture Media, Conditioned; D006493:Heparin