Human umbilical cord blood mesenchymal stem cells expansion via human fibroblast-derived matrix and their potentials toward regenerative application
Large expansion of human mesenchymal stem cells (MSCs) is of great interest for clinical applications. In this study, we examine the feasibility of human fibroblast-derived extracellular matrix (hFDM) as an alternative cell expansion setting. hFDM is obtained from decellularized extracellular matrix (ECM) derived from in vitro cultured human lung fibroblasts. Our study directly compares conventional platforms (tissue culture plastic (TCP), fibronectin (FN)-coated TCP) with hFDM using umbilical cord blood-derived MSCs (UCB-MSCs). Early cell morphology shows a rather rounded shape on TCP but highly elongated morphology on hFDM. Cell proliferation demonstrates that MSCs on hFDM were significantly better compared to the others in both 10 and 2% serum condition. Cell migration assay suggests that cell motility was improved and a cell migration marker CXCR4 was notably up-regulated on hFDM. MSCs differentiation into osteogenic lineage on hFDM was also very effective as examined via gene expression, von Kossa staining and alkaline phosphatase activity. In addition, as the MSCs were expanded on each substrate, transferred to 3D polymer mesh scaffolds and then cultivated for a while, the data found better cell proliferation and more CXCR4 expression with MSCs pre-conditioned on hFDM. Moreover, higher gene expression of stemness and engraftment-related markers was noticed with the hFDM group. Furthermore when UCB-MSCs expanded on TCP or hFDM were injected into emphysema (a lung disease) animal model, the results indicate that MSCs pre-conditioned on hFDM (with 2% serum) retain more advanced therapeutic efficacy on the improvement of emphysema than those on TCP. Current works demonstrate that compared to the conventional platforms, hFDM can be a promising source of cell expansion with a naturally derived biomimetic ECM microenvironment and may find some practical applications in regenerative medicine.
KeywordsExtracellular matrix (ECM) Human fibroblast-derived matrix (hFDM) Umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) MSCs expansion Emphysema
This study was supported by the Korea Health Industry Development Institute (KHIDI) and funded by the Ministry of Health and Welfare (HI16C0133), Republic of Korea.
Compliance with ethical standards
The authors declare that they have no competing financial interests.
- Du P, Subbiah R, Park JH, Park K (2014) Vascular morphogenesis of human umbilical vein endothelial cells on cell-derived macromolecular matrix microenvironment. Tissue Eng A:2365–2377Google Scholar
- Huh JW, Kim SY, Lee JH, Lee JS, Van Ta Q, Kim M, Oh YM, Lee YS, Lee SD (2011) Bone marrow cells repair cigarette smoke-induced emphysema in rats. Am J Phys Lung Cell Mol Phys 301:L255–L266Google Scholar
- Ko JY, Lee J, Lee J, Im GI (2017) Intra-articular xenoplantation of adipose-derived stromal cells to treat osteoarthritis in a goat model. TERM 14:65–71Google Scholar
- Korzyńska A, Zychowicz M (2008) A method of estimation of the cell doubling time on basis of the cell culture monitoring data. BBE 28:75–82Google Scholar
- Kucia M, Ratajczak J, Reca R, Janowska-Wieczorek A, Ratajczak MZ (2004) Tissue-specific muscle, neural and liver stem/progenitor cells reside in the bone marrow, respond to an SDF-1 gradient and are mobilized into peripheral blood during stress and tissue injury. Blood Cells Mol Dis 32:52–57CrossRefGoogle Scholar
- Kusuma GK, Yang MC, Brennecke SP, O’Connor AJ, Kalionis B, Heath DE (2018) Transferable matrixes produced from decellularized extracellular matrix promote proliferation and osteogenic differentiation of mesenchymal stem cells and facilitate scale-up. ACS Biomater Sci Eng. https://doi.org/10.1021/acsbiomaterials.7b00747
- Li O, Tormin A, Sundberg B, Hyllner J, Le Blanc K, Scheding S (2013) Human embryonic stem cell-derived mesenchymal stroma cells (hES-MSCs) engraft in vivo and support hematopoiesis without suppressing immune function: implications for off-the shelf ES-MSC therapies. PLoS One 8:e55319CrossRefGoogle Scholar
- Moriscot C, de Fraipont F, Richard MJ, Marchand M, Savatier P, Bosco D, Favrot M, Benhamou PY (2015) Human bone marrow mesenchymal stem cells can express insulin and key transcription factors of the endocrine pancreas developmental pathway upon genetic and/or microenvironmental manipulation in vitro. Stem Cells 23:594–603CrossRefGoogle Scholar
- Rabe KF, Hurd S, Anzueto A, Barnes PJ, Buist SA, Calverley P, Fukuchi Y, Jenkins C, Rodriguez-Roisin R, van Weel C, Zielinski J (2007) Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary. Am J Respir Crit Care Med 176:532–555CrossRefGoogle Scholar
- Wang Y, Zhang Z, Chi Y, Zhang Q, Xu F, Yang Z, Meng L, Yang S, Yan S, Mao A, Zhang J, Yang Y, Wang S, Cui J, Liang L, Ji Y, Han ZB, Fang X, Han ZC (2013) Long-term cultured mesenchymal stem cells frequently develop genomic mutations but do not undergo malignant transformation. Cell Death Dis 4:e950CrossRefGoogle Scholar