Tissue engineering requires large numbers of cells with enhanced differentiation properties. Thus, the effect of expansion conditions must be explored. Human and rat marrow-derived mesenchymal stem cells (hMSCs and rMSCs, respectively) were comparatively culture expanded through seven passages in the presence of either fibroblast growth factor-2 (FGF-2) or platelet-derived growth factor BB (PDGF-BB). Proliferation of both hMSCs and rMSCs was enhanced by FGF-2 and PDGF-BB. Population doubling times for hMSCs were 2.4 days for control and 1.75 and 2.0 days for FGF-2 and PDGF-BB, respectively, and 3.25, 3.06, and 2.95 days for rMSCs. Supplementation with FGF-2 during cell expansion resulted in significantly greater in vivo bone formation for hMSCs. Use of PDGF-BB resulted in greater bone formation than that observed for control conditions, but the differences were only significant for P1. For rMSCs, significant increases in bone formation were noted in either FGF-2 or PDGF-BB expanded cells implanted at P4 or P7, but not for P1. Under in vitro osteogenic stimulation, calcium content was elevated and bone matrix deposition was enhanced for P1 and P7 rMSCs expanded with FGF-2. Although culture conditions, including FBS, were held constant, these observations suggest that medium must be optimized separately for each species of MSCs.
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References
1.
CaplanA.I., and BruderS.P.Cell and molecular engineering of bone regeneration. In: LanzaR.P., ChickW.L., and LangerR., eds. Principles of Tissue Engineering.New York: R.G. Landes/Springer, 1996, pp. 599–618.
2.
CaplanA.I.Tissue engineering designs for the future: new logics, old molecules. Tissue Eng, 6, 1, 2000.
3.
CaplanA.I.Mesenchymal Stem Cells. In: LanzaR., GearhartJ., HoganB., MeltonD., PedersenR., ThomasE.D., et al., eds. Essentials of Stem Cell Biology. Cambridge, MA: Elsevier Academic Press, 2005, pp. 205–210.
4.
BaiL., LennonD.P., EatonV., et al.Human bone marrow-derived mesenchymal stem cells induce Th2-polarized immune response and promote endogenous repair in animal models of multiple sclerosis. Glia, 57, 1192, 2009.
5.
BeedermanM., LamplotJ.D., NanG., et al.BMP signaling in mesenchymal stem cell differentiation and bone formation. J Biomed Sci Eng, 6, 32, 2013.
6.
NgF., BoucherS., KohS., et al.TGF-beta, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): transcriptional profiling can identify markers and signaling pathways important in differentiation of MSCs into adipogenic, chondrogenic, and osteogenic lineages. Blood, 112, 295, 2008.
7.
XuB., LuoY., LiuY., LiB.Y., and WangY.Platelet-derived growth factor-BB enhances MSC-mediated cardioprotection via suppression of miR-320 expression. Am J Physiol Heart Circ Physiol, 308, H980, 2015.
8.
HanadaK., DennisJ.E., and CaplanA.I.Stimulatory effects of basic fibroblast growth factor and bone morphogenetic protein-2 on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells. J Bone Miner Res, 12, 1606, 1997.
9.
SolchagaL.A., PenickK., GoldbergV.M., CaplanA.I., and WelterJ.F.Fibroblast growth factor-2 enhances proliferation and delays loss of chondrogenic potential in human adult bone-marrow-derived mesenchymal stem cells. Tissue Eng Part A, 16, 1009, 2010.
10.
SolchagaL.A., TemenoffJ.S., GaoJ., MikosA.G., CaplanA.I., and GoldbergV.M.Repair of osteochondral defects with hyaluronan- and polyester-based scaffolds. Osteoarthritis Cartilage, 13, 297, 2005.
11.
HaynesworthS.E., BaberM.A., and CaplanA.I.Cell surface antigens on human marrow-derived mesenchymal cells are detected by monoclonal antibodies. Bone, 13, 69, 1992.
12.
HaynesworthS.E., GoshimaJ., GoldbergV.M., and CaplanA.I.Characterization of cells with osteogenic potential from human marrow. Bone, 13, 81, 1992.
13.
CaplanA.I., and DennisJ.E.Mesenchymal stem cells as trophic mediators. J Cell Biochem, 98, 1076, 2006.
14.
LennonD.P., and CaplanA.I.Isolation of rat marrow-derived mesenchymal stem cells. Exp Hematol, 34, 1606, 2006.
15.
LennonD.P., HaynesworthS.E., BruderS.P., JaiswalN., and CaplanA.I.Human and animal mesenchymal progenitor cells from bone marrow: identification of serum for optimal selection and proliferation. In Vitro Cell Dev Biol Anim, 32, 602, 1996.
16.
OhgushiH., and CaplanA.I.Stem cell technology and bioceramics: from cell to gene engineering. J Biomed Mater Res, 48, 913, 1999.
17.
DennisJ.E., and CaplanA.I.Porous ceramic vehicles for rat-marrow-derived (Rattus norvegicus) osteogenic cell delivery: effects of pre-treatment with fibronectin or laminin. J Oral Implantol, 19, 106, 1993.
18.
GaoJ., DennisJ.E., SolchagaL.A., AwadallahA.S., GoldbergV.M., and CaplanA.I.Tissue-engineered fabrication of an osteochondral composite graft using rat bone marrow-derived mesenchymal stem cells. Tissue Eng, 7, 363, 2001.
19.
LennonD.P., HaynesworthS.E., BruderS.P., JaiswalN., and CaplanA.I.Human and animal mesenchymal progenitor cells from bone marrow: identification of serum for optimal selection and proliferation. In Vitro Cell Dev Anim, 32, 602, 1996.
20.
DennisJ.E., KonstantakosE.K., ArmD., and CaplanA.I.In vivo osteogenesis assay: a rapid method for quantitative analysis. Biomaterials, 19, 1323, 1998.
21.
FriedensteinA.J., ChailakhyanR.K., and GerasimovU.V.Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet, 20, 263, 1987.
22.
JaiswalN., HaynesworthS.E., CaplanA.I., and BruderS.P.Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem, 64, 295, 1997.
23.
NakamuraY., TenshoK., NakayaH., NawataM., OkabeT., and WakitaniS.Low dose fibroblast growth factor-2 (FGF-2) enhances bone morphogenetic protein-2 (BMP-2)-induced ectopic bone formation in mice. Bone, 36, 399, 2005.
24.
Hughes-FulfordM., and LiC.F.The role of FGF-2 and BMP-2 in regulation of gene induction, cell proliferation and mineralization. J Orthop Surg Res, 6, 8, 2011.
25.
ItoT., SawadaR., FujiwaraY., and TsuchiyaT.FGF-2 increases osteogenic and chondrogenic differentiation potentials of human mesenchymal stem cells by inactivation of TGF-beta signaling. Cytotechnology, 56, 1, 2008.
26.
DombrowskiC., SongS.J., ChuanP., et al.Heparan sulfate mediates the proliferation and differentiation of rat mesenchymal stem cells. Stem Cells Dev, 18, 661, 2009.
27.
HuangZ., RenP.G., MaT., SmithR.L., and GoodmanS.B.Modulating osteogenesis of mesenchymal stem cells by modifying growth factor availability. Cytokine, 51, 305, 2010.
28.
CaplanA.I., and CorreaD.PDGF in bone formation and regeneration: new insights into a novel mechanism involving MSCs. J Orthop Res, 29, 1795, 2011.
29.
PountosI., GeorgouliT., HenshawK., BirdH., and GiannoudisP.V.Release of growth factors and the effect of age, sex, and severity of injury after long bone fracture. A preliminary report. Acta Orthop, 84, 65, 2013.
30.
LennonD.P., SchluchterM.D., and CaplanA.I.The effect of extended first passage culture on the proliferation and differentiation of human marrow-derived mesenchymal stem cells. Stem Cells Transl Med, 1, 279, 2012.
