We have previously isolated mesenchymal stromal cells (MSCs) from the tracheal aspirates of premature neonates with respiratory distress. Although isolation of MSCs correlates with the development of bronchopulmonary dysplasia, the physiologic role of these cells remains unclear. To address this, we further characterized the cells, focusing on the issues of gene expression, origin, and cytokine expression. Microarray comparison of early passage neonatal lung MSC gene expression to cord blood MSCs and human fetal and neonatal lung fibroblast lines demonstrated that the neonatal lung MSCs differentially expressed 971 gene probes compared with cord blood MSCs, including the transcription factors Tbx2, Tbx3, Wnt5a, FoxF1, and Gli2, each of which has been associated with lung development. Compared with lung fibroblasts, 710 gene probe transcripts were differentially expressed by the lung MSCs, including IL-6 and IL-8/CXCL8. Differential chemokine expression was confirmed by protein analysis. Further, neonatal lung MSCs exhibited a pattern of Hox gene expression distinct from cord blood MSCs but similar to human fetal lung fibroblasts, consistent with a lung origin. On the other hand, limiting dilution analysis showed that fetal lung fibroblasts form colonies at a significantly lower rate than MSCs, and fibroblasts failed to undergo differentiation along adipogenic, osteogenic, and chondrogenic lineages. In conclusion, MSCs isolated from neonatal tracheal aspirates demonstrate a pattern of lung-specific gene expression, are distinct from lung fibroblasts, and secrete pro-inflammatory cytokines.
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References
1.
JobeAH, BancalariE. 2001. Bronchopulmonary dysplasia. Am J Respir Crit Care Med, 163:1723–1729.
2.
EberE, ZachMS. 2001. Paediatric origins of adult lung disease {bullet} 8: long term sequelae of bronchopulmonary dysplasia (chronic lung disease of infancy)Thorax, 56:317–323.
3.
HussainNA, SiddiquiNH, StockerJR. 1998. Pathology of arrested acinar development in postsurfactant bronchopulmonary dysplasia. Hum Pathol, 29:710–717.
4.
CoalsonJJ. 2003. Pathology of new bronchopulmonary dysplasia. Semin Neonatol, 8:73–81.
5.
BhattAJ, PryhuberGS, HuyckH, WatkinsRH, MetlayLA, ManiscalcoWM. 2001. Disrupted pulmonary vasculature and decreased vascular endothelial growth factor, Flt-1, and TIE-2 in human infants dying with bronchopulmonary dysplasia. Am J Respir Crit Care Med, 164:1971–1980.
6.
TotiP, BuonocoreG, TanganelliP, CatellaAM, PalmeriML, VattiR, SeemayerTA. 1997. Bronchopulmonary dysplasia of the premature baby: an immunohistochemical study. Pediatr Pulmonol, 24:22–28.
7.
HennrickKT, KeetonAG, NanuaS, KijekTG, GoldsmithAM, SajjanUS, BentleyJK, LamaVN, MooreBB, SchumacherRE, ThannickalVJ, HershensonMB. 2007. Lung cells from neonates show a mesenchymal stem cell phenotype. Am J Respir Crit Care Med, 175:1158–1164.
8.
PopovaAP, BozykPD, GoldsmithAM, LinnMJ, LeiJ, BentleyJK, HershensonMB. 2010. Autocrine production of TGF-b1 promotes myofibroblastic differentiation of neonatal lung mesenchymal stem cells. Am J Physiol Lung Cell Mol Physiol, 298:L735–L743.
9.
PopovaAP, BozykPD, BentleyJK, LinnMJ, GoldsmithAM, SchumacherRE, FilbrunAG, WeinerGM, HershensonMB. 2010. Isolation of mesenchymal stromal cells from tracheal aspirates of premature infants predicts bronchopulmonary dysplasia. Pediatrics, 126:e1127–e1133.
10.
SarugaserR, HanounL, KeatingA, StanfordWL, DaviesJE. 2009. Human mesenchymal stem cells self-renew and differentiate according to a deterministic hierarchy. PLoS One, 4:e6498.
da Silva MeirellesL, ChagastellesPC, NardiNB. 2006. Mesenchymal stem cells reside in virtually all post-natal organs and tissues. J Cell Sci, 119:2204–2213.
13.
CrisanM, YapS, CasteillaL, ChenC-W, CorselliM, ParkTS, AndrioloG, SunB, ZhengB, ZhangL, NorotteC, TengP-N, TraasJ, SchugarR, DeasyBM, BadylakS, BuhringH-J, GiacobinoJ-P, LazzariL, HuardJ, PéaultB. 2008. A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell, 3:301–313.
14.
LamaVN, SmithL, BadriL, FlintAJ, AndreiA-C, MurrayS, WangZ, LiaoH, ToewsGB, KrebsbachPH, Peters-GoldenM, PinskyDJ, MartinezFJ, ThannickalVJ. 2007. Evidence for tissue-resident mesenchymal stem cells in human adult lung from studies of transplanted allografts. J Clin Invest, 117:989–996.
15.
SueblinvongV, LoiR, EisenhauerPL, BernsteinIM, SurattBT, SpeesJL, WeissDJ. 2008. Derivation of lung epithelium from human cord blood-derived mesenchymal stem cells. Am J Respir Crit Care Med, 177:701–711.
16.
BenjaminiY, HochbergY. 1995. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc B, 57:289–300.
17.
DennisG, ShermanBT, HosackDA, YangJ, GaoW, LaneHC, LempickiRA. 2003. DAVID: database for annotation, visualization, and integrated discovery. Genome Biol, 4:P3.
18.
HuangDW, ShermanBT, LempickiRA. 2009. Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources. Nat Protoc, 4:44–57.
19.
EhrenkranzRA, WalshMC, VohrBR, JobeAH, WrightLL, FanaroffAA, WrageLA, PooleK. for the National Institutes of Child Health Human Development Neonatal Research Network. 2005. Validation of the National Institutes of Health Consensus Definition of Bronchopulmonary Dysplasia. Pediatrics, 116:1353–1360.
20.
in't AnkerPS, NoortWA, ScherjonSA, Kleijburg-van der KeurC, KruisselbrinkAB, van BezooijenRL, BeekhuizenW, WillemzeR, KanhaiHH, FibbeWE. 2003. Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential. Haematologica, 88:845–852.
21.
ChangHY, ChiJ-T, DudoitS, BondreC, van de RijnM, BotsteinD, BrownPO. 2002. Diversity, topographic differentiation, and positional memory in human fibroblasts. Proc Natl Acad Sci USA, 99:12877–12882.
22.
AckemaKB, ChariteJ. 2008. Mesenchymal stem cells from different organs are characterized by distinct topographic Hox codes. Stem Cells and Development, 17:979–992.
23.
YaekashiwaM, NakayamaS, OhnumaK, SakaiT, AbeT, SatohKEN, MatsumotoK, NakamuraT, TakahashiT, NukiwaT. 1997. Simultaneous or Delayed Administration of Hepatocyte Growth Factor Equally Represses the Fibrotic Changes in Murine Lung Injury Induced by Bleomycin. A Morphologic Study. Am J Respir Crit Care Med, 156:1937–1944.
24.
DohiM, HasegawaT, YamamotoK, MarshallBC. 2000. Hepatocyte Growth Factor Attenuates Collagen Accumulation in a Murine Model of Pulmonary Fibrosis. Am J Respir Crit Care Med, 162:2302–2307.
25.
GazdharA, FachingerP, van LeerC, PierogJ, GuggerM, FriisR, SchmidRA, GeiserT. 2007. Gene transfer of hepatocyte growth factor by electroporation reduces bleomycin-induced lung fibrosis. Am J Physiol Lung Cell Mol Physiol, 292:L529–L536.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD, HorwitzE. 2006. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
28.
HashimotoN, JinH, LiuT, ChensueSW, PhanSH. 2004. Bone marrow-derived progenitor cells in pulmonary fibrosis. J Clin Invest, 113:243–252.
29.
WongAP, KeatingA, LuW-Y, DuchesneauP, WangX, SacherA, HuJ, WaddellTK. 2009. Identification of a bone marrow-derived epithelial-like population capable of repopulating injured mouse airway epithelium. J Clin Invest, 119:336–348.
30.
BoucheratO, Franco-MontoyaML, ThibaultC, IncittiR, Chailley-HeuB, DelacourtC, BourbonJR. 2007. Gene expression profiling in lung fibroblasts reveals new players in alveolarization. Physiol Genomics, 32:128–141.
31.
MandevilleI, AubinJ, LeBlancM, Lalancette-HebertM, JanelleMF, TremblayGM, JeannotteL. 2006. Impact of the loss of Hoxa5 function on lung alveogenesis. Am J Pathol, 169:1312–1327.
32.
VolpeMV, VosatkaRJ, NielsenHC. 2000. Hoxb-5 control of early airway formation during branching morphogenesis in the developing mouse lung. Biochim Biophys Acta, 1475:337–345.
33.
VolpeMV, PhamL, LessinM, RalstonSJ, BhanI, CutzE, NielsenHC. 2003. Expression of Hoxb-5 during human lung development and in congenital lung malformations. Birth Defects Res A Clin Mol Teratol, 67:550–556.
34.
YamaguchiTP, BradleyA, McMahonAP, JonesS. 1999. A Wnt5a pathway underlies outgrowth of multiple structures in the vertebrate embryo. Development, 126:1211–1223.
35.
LiC, XiaoJ, HormiK, BorokZ, MinooP. 2002. Wnt5a participates in distal lung morphogenesis. Dev Biol, 248:68–81.
36.
KalinichenkoVV, LimL, StolzDB, ShinB, RausaFM, ClarkJ, WhitsettJA, WatkinsSC, CostaRH. 2001. Defects in pulmonary vasculature and perinatal lung hemorrhage in mice heterozygous null for the Forkhead Box f1 transcription factor. Dev Biol, 235:489–506.
37.
ChapmanDL, GarveyN, HancockS, AlexiouM, AgulnikSI, Gibson-BrownJJ, Cebra-ThomasJ, BollagRJ, SilverLM, PapaioannouVE. 1996. Expression of the T-box family genes, Tbx1-Tbx5, during early mouse development. Dev Dyn, 206:379–390.
38.
Cebra-ThomasJA, BromerJ, GardnerR, LamGK, SheipeH, GilbertSF. 2003. T-box gene products are required for mesenchymal induction of epithelial branching in the embryonic mouse lung. Dev Dyn, 226:82–90.
39.
JarvinenL, BadriL, WettlauferS, OhtsukaT, StandifordTJ, ToewsGB, PinskyDJ, Peters-GoldenM, LamaVN. 2008. Lung resident mesenchymal stem cells isolated from human lung allografts inhibit T cell proliferation via a soluble mediator. J Immunol, 181:4389–4396.
40.
BesnardV, WertSE, StahlmanMT, PostleAD, XuY, IkegamiM, WhitsettJA. 2009. Deletion of Scap in alveolar type II cells influences lung lipid homeostasis and identifies a compensatory role for pulmonary lipofibroblasts. J Biol Chem, 284:4018–4030.
41.
McGowanSE, TordayJS. 1997. The pulmonary lipofibroblast (lipid interstitial cell) and its contributions to alveolar development-1. Ann Rev Physiol, 59:43–62.
42.
BoströmH, WillettsK, PeknyM, LevéenP, LindahlP, HedstrandH, PeknaM, HellströmM, Gebre-MedhinS, SchallingM, NilssonM, KurlandS, TörnellJ, HeathJK, BetsholtzC. 1996. PDGF-A signaling is a critical event in lung alveolar myofibroblast development and alveogenesis. Cell, 85:863–873.
43.
LindahlP, KarlssonL, HellstromM, Gebre-MedhinS, WillettsK, HeathJ, BetsholtzC. 1997. Alveogenesis failure in PDGF-A-deficient mice is coupled to lack of distal spreading of alveolar smooth muscle cell progenitors during lung development. Development, 124:3943–3953.
44.
RehanVK, SuganoS, WangY, SantosJ, RomeroS, DasguptaC. 2006. Evidence for the presence of lipofibroblasts in human lung. Exp Lung Res, 32:379–393.
45.
VadivelA, AbozaidS, van HaaftenT, SawickaM, EatonF, ChenM, ThebaudB. 2010. Adrenomedullin promotes lung angiogenesis, alveolar development and repair. Am J Respir Cell Mol Biol, 43:152–160.
ScottonCJ, KrupiczojcMA, KonigshoffM, MercerPF, LeeYCG, KaminskiN, MorserJ, PostJM, MaherTM, NicholsonAG, MoffattJD, LaurentGJ, DerianCK, EickelbergO, ChambersRC. 2009. Increased local expression of coagulation factor X contributes to the fibrotic response in human and murine lung injury. J Clin Invest, 119:2550–2563.
48.
van HaaftenT, ByrneR, BonnetS, RochefortGY, AkabutuJ, BouchentoufM, Rey-ParraGJ, GalipeauJ, HaromyA, EatonF, ChenM, HashimotoK, AbleyD, KorbuttG, ArcherSL, ThebaudB. 2009. Airway delivery of mesenchymal stem cells prevents arrested alveolar growth in neonatal lung injury in rats. Am J Respir Crit Care Med, 180:1131–1142.
49.
AslamM, BavejaR, LiangOD, Fernandez-GonzalezA, LeeC, MitsialisSA, KourembanasS. 2009. Bone marrow stromal cells attenuate lung injury in a murine model of neonatal chronic lung disease. Am J Respir Crit Care Med, 180:1122–1130.
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