Iranian Journal of Allergy, Asthma and Immunology 2017. 16(6):501-510.

Attenuating Effect of Long-term Culture of Umbilical Cord Vein Mesenchymal Stromal Cells on Pulmonary Fibrosis in C57BL/6 Mice
Maryam Moradi, Mohammad Ali Rezaee, Mehdi Mohammadi, Mohammad Jafar Rezaie, Ali Jalili, Mohammad Reza Rahmani


In recent studies, mesenchymal stromal cells (MSCs) have been increasingly employed to treat various diseases like pulmonary fibrosis (PF). There are very few MSCs in tissues so in order to obtain their sufficient numbers for therapeutic applications, their in vitro expansion is necessary. The aim of this study was to investigate the effects of long-term culture of the human umbilical cord vein MSCs (hUCV-MSCs) on pulmonary fibrosis in mice. MSCs were first isolated from human umbilical cord vein and cultured up to 18 passages. In C57BL/6 mice, 15 min after belomycin instillation, UCV-MSCs at passages (P) 0, 4, 8, 12, and 18 (long-term culture) were transplanted intratracheally. Mice were weighted every 5 days and were euthanized on day 21. For histopathological examination, the lung sections were stained with hematoxylin-eosin (HE) and Masson’s trichrome. The mRNA expression of TGF-β1, alpha-smooth muscle actin (α-SMA), and collagen type I alpha 1 (COL1A1) in lung tissues were assessed using RT-PCR. For cell tracking, human cytochrome B DNA was detected in mice lung tissues by PCR. The weight of mice receiving long-term culture of UCV-MSCs increased compared to other mice (p=0.056). Also, transplantation of UCV-MSCs at P18 led to increased alveolar space and decreased connective tissue and collagen deposition of the lung tissues. The mRNA expression of TGF-β1, α-SMA, and COL1A1 also decreased in this group. The results showed that intratracheally transplanted long-term culture of the UCV-MSCs attenuated lung fibrosis in mice. 


Bleomycin; Long-term culture; Mesenchymal stromal cell (MSC); Pulmonary fibrosis

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[1.           Wynn TA, Ramalingam TR. Mechanisms of fibrosis: therapeutic translation for fibrotic disease. Nature medicine. 2012;18(7):1028-40.

2.            Ariel A, Timor O. Hanging in the balance: endogenous anti‐inflammatory mechanisms in tissue repair and fibrosis. The Journal of pathology. 2013;229(2):250-63.

3.            Cheresh P, Kim S-J, Tulasiram S, Kamp DW. Oxidative stress and pulmonary fibrosis. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease. 2013;1832(7):1028-40.

4.            Kalayarasan S, Sriram N, Sudhandiran G. Diallyl sulfide attenuates bleomycin-induced pulmonary fibrosis: Critical role of iNOS, NF-κB, TNF-α and IL-1β. Life sciences. 2008;82(23):1142-53.

5.            Lim MJ, Ahn J, Yi JY, Kim M-H, Son A-R, Lim D-S, et al. Induction of galectin-1 by TGF-β1 accelerates fibrosis through enhancing nuclear retention of Smad2. Experimental cell research. 2014;326(1):125-35.

6.            Kang Y-Y, Kim D-Y, Lee S-H, Choi EY. Deficiency of developmental endothelial locus-1 (Del-1) aggravates bleomycin-induced pulmonary fibrosis in mice. Biochemical and biophysical research communications. 2014;445(2):369-74.

7.            Si G, Tao Z, Wei W, Min X, Wang X-c, Chen Z-h. Glucagon like peptide-1 attenuates bleomycin-induced pulmonary fibrosis, involving the inactivation of NF-κB in mice. International immunopharmacology. 2014;22(2):498-504.

8.            Datta A, Scotton CJ, Chambers RC. Novel therapeutic approaches for pulmonary fibrosis. British journal of pharmacology. 2011;163(1):141-72.

9.            Wang Z, Zhang X, Kang Y, Zeng Y, Liu H, Chen X, et al. Stem cell therapy for idiopathic pulmonary fibrosis: How far are we from the bench to the bedside? Journal of Biomedical Science and Engineering. 2013;2013.

10.          Toonkel RL, Hare JM, Matthay MA, Glassberg MK. Mesenchymal stem cells and idiopathic pulmonary fibrosis. Potential for clinical testing. American journal of respiratory and critical care medicine. 2013;188(2):133-40.

11.          Xu J, Mora A, Shim H, Stecenko A, Brigham KL, Rojas M. Role of the SDF-1/CXCR4 axis in the pathogenesis of lung injury and fibrosis. American journal of respiratory cell and molecular biology. 2007;37(3):291-9.

12.          Moodley Y, Atienza D, Manuelpillai U, Samuel CS, Tchongue J, Ilancheran S, et al. Human umbilical cord mesenchymal stem cells reduce fibrosis of bleomycin-induced lung injury. The American journal of pathology. 2009;175(1):303-13.

13.          Zhao F, Zhang Y, Liu Y, Zhou J, Li Z, Wu C, et al., editors. Therapeutic effects of bone marrow-derived mesenchymal stem cells engraftment on bleomycin-induced lung injury in rats. Transplantation proceedings; 2008: Elsevier.

14.          Cargnoni A, Gibelli L, Tosini A, Signoroni PB, Nassuato C, Arienti D, et al. Transplantation of allogeneic and xenogeneic placenta-derived cells reduces bleomycin-induced lung fibrosis. Cell transplantation. 2009;18(4):405-22.

15.          Wexler SA, Donaldson C, Denning-Kendall P, Rice C, Bradley B, Hows JM. Adult bone marrow is a rich source of human mesenchymal 'stem' cells but umbilical cord and mobilized adult blood are not. British journal of haematology. 2003;121(2):368-74.

16.          Choi MR, Kim HY, Park J-Y, Lee TY, Baik CS, Chai YG, et al. Selection of optimal passage of bone marrow-derived mesenchymal stem cells for stem cell therapy in patients with amyotrophic lateral sclerosis. Neuroscience letters. 2010;472(2):94-8.

17.          Bonab MM, Alimoghaddam K, Talebian F, Ghaffari SH, Ghavamzadeh A, Nikbin B. Aging of mesenchymal stem cell in vitro. BMC cell biology. 2006;7(1):1.

18.          Wagner W, Bork S, Lepperdinger G, Joussen S, Ma N, Strunk D, et al. How to track cellular aging of mesenchymal stromal cells. Aging (Albany NY). 2010;2(4):224-30.

19.          Kim J, Kang JW, Park JH, Choi Y, Choi KS, Park KD, et al. Biological characterization of long-term cultured human mesenchymal stem cells. Archives of pharmacal research. 2009;32(1):117-26.

20.          Zhuang Y, Li D, Fu J, Shi Q, Lu Y, Ju X. Comparison of biological properties of umbilical cord‑derived mesenchymal stem cells from early and late passages: Immunomodulatory ability is enhanced in aged cells. Molecular medicine reports. 2015;11(1):166-74.

21.          Song D, Ohtaki H, Tsumuraya T, Miyamoto K, Shibato J, Rakwal R, et al. The anti-inflammatory property of human bone marrow-derived mesenchymal stem/stromal cells is preserved in late-passage cultures. Journal of neuroimmunology. 2013;263(1):55-63.

22.          Li S, Wang Y, Guan L, Ji M. Characteristics of human umbilical cord mesenchymal stem cells during ex vivo expansion. Molecular medicine reports. 2015;12(3):4320-5.

23.          Nur Fariha M-M, Chua K-H, Tan G-C, Tan A-E, Hayati A-R. Human chorion-derived stem cells: changes in stem cell properties during serial passage. Cytotherapy. 2011;13(5):582-93.

24.          Kadivar M, Khatami S, Mortazavi Y, Soleimani M, Taghikhani M, Shokrgozar MA. Isolation, culture and characterization of postnatal human umbilical vein-derived mesenchymal stem cells. DARU Journal of Pharmaceutical Sciences. 2005;13(4):170-6.

25.          Gharaee-Kermani M, Ullenbruch M, Phan SH. Animal models of pulmonary fibrosis.  Fibrosis Research: Springer; 2005. p. 251-9.

26.          Nalysnyk L, Cid-Ruzafa J, Rotella P, Esser D. Incidence and prevalence of idiopathic pulmonary fibrosis: review of the literature. European Respiratory Review. 2012;21(126):355-61.

27.          Gharaee-Kermani M, Gyetko MR, Hu B, Phan SH. New insights into the pathogenesis and treatment of idiopathic pulmonary fibrosis: a potential role for stem cells in the lung parenchyma and implications for therapy. Pharmaceutical research. 2007;24(5):819-41.

28.          Xu Y, Liu Y, Wang Q, Li G, Lü X, Kong B. Intravenous transplantation of mesenchymal stem cells attenuates oleic acid induced acute lung injury in rats. Chinese medical journal. 2012;125(11).

29.          Gupta N, Su X, Popov B, Lee JW, Serikov V, Matthay MA. Intrapulmonary delivery of bone marrow-derived mesenchymal stem cells improves survival and attenuates endotoxin-induced acute lung injury in mice. The Journal of Immunology. 2007;179(3):1855-63.

30.          Tashiro J, Elliot SJ, Gerth DJ, Xia X, Pereira-Simon S, Choi R, et al. Therapeutic benefits of young, but not old, adipose-derived mesenchymal stem cells in a chronic mouse model of bleomycin-induced pulmonary fibrosis. Translational Research. 2015;166(6):554-67.

31.          Chang YS, Oh W, Choi SJ, Sung DK, Kim SY, Choi EY, et al. Human umbilical cord blood-derived mesenchymal stem cells attenuate hyperoxia-induced lung injury in neonatal rats. Cell transplantation. 2009;18(8):869-86.

32.          Izadpanah R, Kaushal D, Kriedt C, Tsien F, Patel B, Dufour J, et al. Long-term in vitro expansion alters the biology of adult mesenchymal stem cells. Cancer research. 2008;68(11):4229-38.

33.          Kuwano K, Nakashima N, Inoshima I, Hagimoto N, Fujita M, Yoshimi M, et al. Oxidative stress in lung epithelial cells from patients with idiopathic interstitial pneumonias. European Respiratory Journal. 2003;21(2):232-40.

34.          Luzina IG, Todd NW, Sundararajan S, Atamas SP. The cytokines of pulmonary fibrosis: Much learned, much more to learn. Cytokine. 2015;74(1):88-100.

35.          Nagamoto T, Eguchi G, Beebe DC. Alpha-smooth muscle actin expression in cultured lens epithelial cells. Investigative ophthalmology & visual science. 2000;41(5):1122-9.


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