Native extracellular matrix preserves mesenchymal stem cell "stemness" and differentiation potential under serum-free culture conditions

被引:59
作者
Rakian, Rubie [1 ]
Block, Travis J. [1 ]
Johnson, Shannan M. [1 ,2 ]
Marinkovic, Milos [1 ]
Wu, Junjie [3 ]
Dai, Qiuxia [1 ]
Dean, David D. [1 ]
Chen, Xiao-Dong [1 ,4 ]
机构
[1] Univ Texas Hlth Sci Ctr San Antonio, Dept Comprehens Dent, San Antonio, TX 78229 USA
[2] Joint Base San Antonio, Periodont Grad Program, Lackland AFB, TX 78236 USA
[3] Fourth Mil Med Univ, Sch Stomatol, Dept Orthodont, Xian 710032, Shaanxi Provinc, Peoples R China
[4] South Texas Vet Hlth Care Syst, Res Serv, Audie Murphy VA Med Ctr, San Antonio, TX 78229 USA
关键词
Mesenchymal stem cells; Extracellular matrix; Serum-free media; Stem cell expansion; MARROW STROMAL CELLS; BONE-MARROW; OSTEOBLAST DIFFERENTIATION; PROGENITOR CELLS; EXPANSION; TRANSPLANTATION; PROLIFERATION; PROTEOGLYCANS; STABILITY; MIGRATION;
D O I
10.1186/s13287-015-0235-6
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Introduction: Bone marrow-derived mesenchymal stem cells (BM-MSCs) for clinical use should not be grown in media containing fetal bovine serum (FBS), because of serum-related concerns over biosafety and batch-to-batch variability. Previously, we described the preparation and use of a cell-free native extracellular matrix (ECM) made by bone marrow cells (BM-ECM) which preserves stem cell properties and enhances proliferation. Here, we compare colony-forming ability and differentiation of MSCs cultured on BM-ECM with a commercially available matrix (CELLstart (TM)) and tissue culture plastic (TCP) under serum-free conditions. Methods: Primary MSCs from freshly isolated bone marrow-derived mononuclear cells or passaged MSCs (P1) were grown in serum-containing (SCM) or serum-free (SFM) media on BM-ECM, CELLstart (TM), or TCP substrates. Proliferation, cell composition (phenotype), colony-forming unit replication, and bone morphogenetic protein-2 (BMP-2) responsiveness were compared among cells maintained on the three substrates. Results: Proliferation of primary BM-MSCs was significantly higher in SCM than SFM, irrespectively of culture substrate, suggesting that the expansion of these cells requires SCM. In contrast, passaged cells cultured on BM-ECM or CELLstart (TM) in SFM proliferated to nearly the same extent as cells in SCM. However, morphologically, those on BM-ECM were smaller and more aligned, slender, and long. Cells grown for 7 days on BM-ECM in SFM were 20-40 % more positive for MSC surface markers than cells cultured on CELLstart (TM). Cells cultured on TCP contained the smallest number of cells positive for MSC markers. MSC colony-forming ability in SFM, as measured by CFU-fibroblasts, was increased 10-, 9-, and 2-fold when P1 cells were cultured on BM-ECM, CELLstart (TM), and TCP, respectively. Significantly, CFU-adipocyte and -osteoblast replication of cells grown on BM-ECM was dramatically increased over those on CELLstart (TM) (2X) and TCP (4-7X). BM-MSCs, cultured in SFM and treated with BMP-2, retained their differentiation capacity better on BM-ECM than on either of the other two substrates. Conclusions: Our findings indicate that BM-ECM provides a unique microenvironment that supports the colony-forming ability of MSCs in SFM and preserves their stem cell properties. The establishment of a robust culture system, combining native tissue-specific ECM and SFM, provides an avenue for preparing significant numbers of potent MSCs for cell-based therapies in patients.
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页数:11
相关论文
共 37 条
[1]   Cell culture: Biology's new dimension [J].
Abbott, A .
NATURE, 2003, 424 (6951) :870-872
[2]   Feasibility and efficacy of bone tissue engineering using human bone marrow stromal cells cultivated in serum-free conditions [J].
Agata, Hideki ;
Watanabe, Nobukazu ;
Ishii, Yumiko ;
Kubo, Noriyuki ;
Ohshima, Satoshi ;
Yamazaki, Mika ;
Tojo, Arinobu ;
Kagami, Hideaki .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2009, 382 (02) :353-358
[3]   Self-Assembled Extracellular Macromolecular Matrices and Their Different Osteogenic Potential with Preosteoblasts and Rat Bone Marrow Mesenchymal Stromal Cells [J].
Bae, Soon Eon ;
Bhang, Suk Ho ;
Kim, Byung-Soo ;
Park, Kwideok .
BIOMACROMOLECULES, 2012, 13 (09) :2811-2820
[4]   Characterization of bone marrow-derived mesenchymal stem cells in aging [J].
Baker, Natasha ;
Boyette, Lisa B. ;
Tuan, Rocky S. .
BONE, 2015, 70 :37-47
[5]   Concise Review: Bone Marrow-Derived Mesenchymal Stem Cells Change Phenotype Following In Vitro Culture: Implications for Basic Research and the Clinic [J].
Bara, Jennifer J. ;
Richards, R. Geoff ;
Alini, Mauro ;
Stoddart, Martin J. .
STEM CELLS, 2014, 32 (07) :1713-1723
[6]   Extracellular matrix proteoglycans control the fate of bone marrow stromal cells [J].
Bi, YM ;
Stuelten, CH ;
Kilts, T ;
Wadhwa, S ;
Iozzo, RV ;
Robey, PG ;
Chen, XD ;
Young, MF .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (34) :30481-30489
[7]  
Brunner D, 2010, ALTEX-ALTERN ANIM EX, V27, P53
[8]   A novel serum-free medium for the expansion of human mesenchymal stem cells [J].
Chase, Lucas G. ;
Lakshmipathy, Uma ;
Solchaga, Luis A. ;
Rao, Mahendra S. ;
Vemuri, Mohan C. .
STEM CELL RESEARCH & THERAPY, 2010, 1
[9]   The small leucine-rich proteoglycan biglycan modulates BMP-4-induced osteoblast differentiation [J].
Chen, XD ;
Fisher, LW ;
Robey, PG ;
Young, MF .
FASEB JOURNAL, 2004, 18 (09) :948-958
[10]   Extracellular matrix made by bone marrow cells facilitates expansion of marrow-derived mesenchymal progenitor cells and prevents their differentiation into osteoblasts [J].
Chen, Xiao-Dong ;
Dusevich, Vladimir ;
Feng, Jian Q. ;
Manolagas, Stavros C. ;
Jilka, Robert L. .
JOURNAL OF BONE AND MINERAL RESEARCH, 2007, 22 (12) :1943-1956