Human mesenchymal stem cells (hMSCs) are promising feeder cells for expanding hematopoietic stem cells (HSCs), but their potential is heterogeneous. We examined the hematopoiesis-supporting activity of hMSC at the clonal level in relation to the osteogenic potential and gene expression. Hematopoiesis-supporting activities of stably immortalized clonal hMSC lines were evaluated by the expansion of CD34+CD38- cells after 7-day coculture with human cord blood-derived CD34+ cells. Six of 16 clones expanded the numbers of CD34+CD38- cells >500-fold. These hematopoiesis-supportive clones also showed high gene expression of Jagged1, a Notch ligand, as well as high potential to deposit calcium after osteogenic induction. Thus, osteogenic hMSC clones may provide proper microenvironments for HSC expansion, ultimately conveying self-renewal signals to HSCs via the Notch pathway. However, they lost hematopoiesis-supporting activity after osteogenic differentiation. The hematopoiesis-supportive clones are potentially useful for hematopoietic microenvironment studies and as components of a coculture system for expansion of HSCs, free from contamination by xenogeneic pathogens.
AdamsG. B.ChabnerK. T.AlleyI. R.OlsonD. P.SzczepiorkowskiZ. M.PoznanskyM. C.KosC. H.PollakM. R.BrownE. M.ScaddenD. T.Stem cell engraftment at the endosteal niche is specified by the calcium-sensing receptor.Nature439: 599–603; 2006.
2.
AmosT. A. S.GordonM. Y.Sources of human hematopoietic stem cells for transplantation—a review.Cell Transplant.4: 547–569; 1995.
3.
Androutsellis-TheotokisA.LekerR. R.SoldnerF.HoeppnerD. J.RavinR.PoserS. W.RuegerM. A.BaeS. K.KittappaR.McKayR. D. G.Notch signalling regulates stem cell numbers in vitro and in vivo.Nature442: 823–826; 2006.
4.
AraiF.HiraoA.OhmuraM.SatoH.MatsuokaS.TakuboK.ItoK.KohG.SudaT.Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche.Cell118: 149–161; 2004.
5.
BreemsD. A.BloklandE. A. W.NebenS.PloemacherR. E.Frequency analysis of human primitive haematopoietic stem cell subsets using a cobblestone area forming cell assay.Leukemia8: 1095–1104; 1994.
DigirolamoC. M.StokesD.ColterD.PhinneyD. G.ClassR.ProckopD. J.Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate.Br. J. Haematol.107: 275–281; 1999.
8.
DucyP.ZhangR.GeoffroyV.RidallA. L.KarsentyG.Osf2/Cbfa1: A transcriptional activator of osteoblast differentiation.Cell89: 747–754; 1997.
9.
FlynnD. M.NijjarS.HubscherS. G.de GoyetJ. D.KellyD. A.StrainA. J.CrosbyH. A.The role of Notch receptor expression in bile duct development and disease.J. Pathol.204: 55–64; 2004.
10.
FujimotoN.FujitaS.TsujiT.ToguchidaJ.IdaK.SuginamiH.IwataH.Microencapsulated feeder cells as a source of soluble factors for expansion of CD34(+) hematopoietic stem cells.Biomaterials28: 4795–4805; 2007.
11.
GronthosS.ChenS.WangC. Y.RobeyP. G.ShiS.Telomerase accelerates osteogenesis of bone marrow stromal stem cells by upregulation of CBFA1, osterix, and osteocalcin.J. Bone Miner. Res.18: 716–722; 2003.
12.
HaoQ. L.ShahA. J.ThiemannF. T.SmogorzewskaE. M.CrooksG. M.A functional comparison of CD34+CD38- cells in cord blood and bone marrow.Blood86: 3745–3753; 1995.
13.
HayE.LemonnierJ.ModrowskiD.LomriA.Las-molesF.MarieP. J.N- and E-cadherin mediate early human calvaria osteoblast differentiation promoted by bone morphogenetic protein-2.J. Cell. Physiol.183: 117–128; 2000.
14.
HofmeisterC. C.ZhangJ.KnightK. L.LeP.StiffP. J.Ex vivo expansion of umbilical cord blood stem cells for transplantation: Growing knowledge from the hematopoietic niche.Bone Marrow Transplant.39: 11–23; 2007.
15.
HuangY. Q.LiJ. J.KarpatkinS.Identification of a family of alternatively spliced mRNA species of angiopoietin-1.Blood95: 1993–1999; 2000.
16.
in't AnkerP. S.NoortW. A.ScherjonS. A.Kleijburg-van der KeurC.KruisselbrinkA. B.van BezooijenR. L.BeekhuizenW.WillemzeR.KanhaiH. H.FibbeW. E.Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential.Haematologica88: 845–852; 2003.
17.
JavazonE. H.ColterD. C.SchwarzE. J.ProckopD. J.Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells.Stem Cells19: 219–225; 2001.
18.
KadereitS.DeedsL. S.HaynesworthS. E.KocO. N.KozikM. M.SzekelyE.Daum-WoodsK.GoetchiusG. W.FuP.WelniakL. A.MurphyW. J.LaughlinM. J.Expansion of LTC-ICs and maintenance of p21 and BCL-2 expression in cord blood CD34(+)/CD38(-) early progenitors cultured over human MSCs as a feeder layer.Stem Cells20: 573–582; 2002.
19.
KawanoY.KobuneM.YamaguchiM.NakamuraK.ItoY.SasakiK.TakahashiS.NakamuraT.ChibaH.SatoT.MatsunagaT.AzumaH.IkebuchiK.IkedaH.KatoJ.NiitsuY.HamadaH.Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells.Blood101: 532–540; 2003.
20.
KiyonoT.FosterS. A.KoopJ. I.McDougallJ. K.GallowayD. A.KlingelhutzA. J.Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells.Nature396: 84–88; 1998.
21.
KuciaM.RatajczakJ.RatajczakM. Z.Are bone marrow stem cells plastic or heterogenous—that is the question.Exp. Hematol.33: 613–623; 2005.
22.
MajumdarM. K.ThiedeM. A.HaynesworthS. E.BruderS. P.GersonS. L.Human marrow-derived mesenchymal stem cells (MSCs) express hematopoietic cytokines and support long-term hematopoiesis when differentiated toward stromal and osteogenic lineages.J. Hematother. Stem Cell Res.9: 841–848; 2000.
23.
ManciniS. J. C.ManteiN.DumortierA.SuterU.MacDonaldH. R.RadtkeF.Jagged1-dependent Notch signaling is dispensable for hematopoietic stem cell self-renewal and differentiation.Blood105: 2340–2342; 2005.
24.
MilonaM. A.GoughJ. E.EdgarA. J.Expression of alternatively spliced isoforms of human Sp7 in osteoblastlike cells.BMC Genomics4: 43; 2003.
25.
NakashimaK.ZhouX.KunkelG.ZhangZ.DengJ. M.BehringerR. R.De CrombruggheB.The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation.Cell108: 17–29; 2002.
26.
Nandoe TewarieR. D. S.HurtadoA.LeviA. D. O.GrotenhuisJ. A.OudegaM.Bone marrow stromal cells for repair of the spinal cord: Towards clinical application.Cell Transplant.15: 563–577; 2006.
27.
NoortW. A.KruisselbrinkA. B.in't AnkerP. S.KrugerM.van BezooijenR. L.de PausR. A.HeemskerkM. H.LöwikC. W.FalkenburgJ. H.WillemzeR.FibbeW. E.Mesenchymal stem cells promote engraftment of human umbilical cord blood-derived CD34(+) cells in NOD/SCID mice.Exp. Hematol.30: 870–878; 2002.
28.
OkamotoT.AoyamaT.NakayamaT.NakamataT.HosakaT.NishijoK.NakamuraT.KiyonoT.ToguchidaJ.Clonal heterogeneity in differentiation potential of immortalized human mesenchymal stem cells.Biochem. Biophys. Res. Commun.295: 354–361; 2002.
29.
PittengerM. F.MackayA. M.BeckS. C.JaiswalR. K.DouglasR.MoscaJ. D.MoormanM. A.SimonettiD. W.CraigS.MarshakD. R.Multilineage potential of adult human mesenchymal stem cells.Science284: 143–147; 1999.
30.
PloemacherR. E.Van Der SluijsJ. P.VoermanJ. S. A.BronsN. H. C.An in vitro limiting-dilution assay of long-term repopulating hematopoietic stem cells in the mouse.Blood74: 2755–2763; 1989.
31.
ReemsJ. A.Torok-StorbB.Cell cycle and functional differences between CD34+/CD38hi and CD34+/38lo human marrow cells after in vitro cytokine exposure.Blood85: 1480–1487; 1995.
32.
ReyaT.DuncanA. W.AillesL.DomenJ.SchererD. C.WillertK.HintzL.NusseR.WeissmanI. L.A role for Wnt signalling in self-renewal of haematopoietic stem cells.Nature423: 409–414; 2003.
33.
RochaV.GluckmanE.Clinical use of umbilical cord blood hematopoietic stem cells.Biol. Blood Marrow Transplant.12: 34–41; 2006.
34.
SutherlandH. J.EavesC. J.LansdorpP. M.ThackerJ. D.HoggeD. E.Differential regulation of primitive human hematopoietic cells in long-term cultures maintained on genetically engineered murine stromal cells.Blood78: 666–672; 1991.
35.
TeraiM.UyamaT.SugikiT.LiX. K.UmezawaA.KiyonoT.Immortalization of human fetal cells: The life span of umbilical cord blood-derived cells can be prolonged without manipulating p16INK4a/RB braking pathway.Mol. Biol. Cell16: 1491–1499; 2005.
36.
TsujiT.OgasawaraH.AokiY.TsurumakiY.KodamaH.Characterization of murine stromal cell clones established from bone marrow and spleen.Leukemia10: 803–812; 1996.
37.
TulacS.NayakN. R.KaoL. C.Van WaesM.HuangJ.LoboS.GermeyerA.LesseyB. A.TaylorR. N.SuchanekE.GiudiceL. C.Identification, characterization, and regulation of the canonical Wnt signaling pathway in human endometrium.J. Clin. Endocrinol. Metab.88: 3860–3866; 2003.
38.
WillertK.BrownJ. D.DanenbergE.DuncanA. W.WeissmanI. L.ReyaT.YatesJ. R.3rdNusseR.Wnt proteins are lipid-modified and can act as stem cell growth factors.Nature423: 448–452; 2003.
39.
ZhangJ.NiuC.YeL.HuangH.HeX.TongW. G.RossJ.HaugJ.JohnsonT.FengJ. Q.HarrisS.WiedemannL. M.MishinaY.LiL.Identification of the haematopoietic stem cell niche and control of the niche size.Nature425: 836–841; 2003.
40.
ZhangY.LiC.JiangX.ZhangS.WuY.LiuB.TangP.MaoN.Human placenta-derived mesenchymal progenitor cells support culture expansion of long-term culture-initiating cells from cord blood CD34+ cells.Exp. Hematol.32: 657–664; 2004.
