Migration of culture-expanded human mesenchymal stem cells through bone marrow endothelium is regulated by matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3

被引:185
作者
De Becker, Ann [1 ]
Van Hummelen, Paul [3 ]
Bakkus, Marleen [2 ]
Vande Broek, Isabelle [1 ]
De Wever, Joke [1 ]
De Waele, Marc [2 ]
Van Riet, Ivan [1 ]
机构
[1] Acad Hosp Vrije Univ Brussel VUB, Stem Cell Lab, Brussels, Belgium
[2] Acad Hosp Vrije Univ Brussel VUB, Lab Hematol Immunol, Brussels, Belgium
[3] Flanders Interuniv Inst Biotechnol VIB, MicroArray Facil, Leuven, Belgium
关键词
mesenchymal stem cell; migration; culture confluence; MMP-2; TIMP-3;
D O I
10.3324/haematol.10475
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Background and Objectives Mesenchymal stem cells (MSC) are adult stem cells that can be expanded many fold in vitro and have the therapeutic potential to restore the bone marrow microenvironment and support hematopoietic recovery after myeloablative conditioning for hematopoietic stem cell transplantation. Successful homing to the target tissue, such as bone marrow, implies that MSC are able to extravasate after systemic administration. However, the extravasation capacity of MSC and the underlying mechanisms are poorly understood to date. We studied in vitro the capacity of MSC to migrate through bone marrow endothelium. Design and Methods In vitro invasion and transendothelial migration assays were performed. The expression of matrix metalloproteinase (MMP) was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Migration of cells cultured at high or low confluence was compared and differential gene expression in these conditions was analyzed with microarray and real-time RT-PCR. The functional involvement in MSC migration was assessed using neutralizing anti-MMP-2 antibody, MMP-2 short interfering RNA or recombinant tissue inhibitor of metalloproteinase (TIMP-3). Results We demonstrated that MSC can invade reconstituted basement membrane and that bone marrow endothelial cells stimulate this process. We also showed that the transendothelial migration of MSC is at least partially regulated by MMP-2. High culture confluence was found to increase production of the natural MMP-inhibitor TIMP-3 and decrease transendothelial migration of MSC. Interpretation and Conclusions We show that MSC have the potential to migrate through bone marrow endothelium and that this process involves MMP-2. Moreover, the migration of MSC is significantly influenced by the level of culture confluence. Increased culture confluence impairs migration and is related to an upregulation of TIMP-3. The therapeutic use of MSC would benefit from a selection of culture conditions that allow optimal extravasation of these cells.
引用
收藏
页码:440 / 449
页数:10
相关论文
共 56 条
[1]   Reconstruction of mandibular defects with autologous tissue-engineered bone [J].
Abukawa, H ;
Shin, M ;
Williams, WB ;
Vacanti, JP ;
Kaban, LB ;
Troulis, MJ .
JOURNAL OF ORAL AND MAXILLOFACIAL SURGERY, 2004, 62 (05) :601-606
[2]   Dynamic of distribution of human bone marrow-derived mesenchymal stem cells after transplantation into adult unconditioned mice [J].
Allers, C ;
Sierralta, WD ;
Neubauer, S ;
Rivera, F ;
Minguell, JJ ;
Conget, PA .
TRANSPLANTATION, 2004, 78 (04) :503-508
[3]   Cotransplantation of human mesenchymal stem cells enhances human myelopoiesis and megakaryocytopoiesis in NOD/SCID mice [J].
Angelopoulou, M ;
Novelli, E ;
Grove, JE ;
Rinder, HM ;
Civin, C ;
Cheng, LZ ;
Krause, DS .
EXPERIMENTAL HEMATOLOGY, 2003, 31 (05) :413-420
[4]   THE GENE STRUCTURE OF TISSUE INHIBITOR OF METALLOPROTEINASES (TIMP)-3 AND ITS INHIBITORY ACTIVITIES DEFINE THE DISTINCT TIMP GENE FAMILY [J].
APTE, SS ;
OLSEN, BR ;
MURPHY, G .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (24) :14313-14318
[5]   Metalloproteinase inhibitors: biological actions and therapeutic opportunities [J].
Baker, AH ;
Edwards, DR ;
Murphy, G .
JOURNAL OF CELL SCIENCE, 2002, 115 (19) :3719-3727
[6]   Retrovirally mediated correction of bone marrow-derived mesenchymal stem cells from patients with mucopolysaccharidosis type I [J].
Baxter, MA ;
Wynn, RF ;
Deakin, JA ;
Bellantuono, I ;
Edington, KG ;
Cooper, A ;
Besley, GTN ;
Church, HJ ;
Wraith, JE ;
Carr, TF ;
Fairbairn, LJ .
BLOOD, 2002, 99 (05) :1857-1859
[7]   Homing of in vitro expanded Stro-1- or Stro-1+ human mesenchymal stem cells into the NOD/SCID mouse and their role in supporting human CD34 cell engraftment [J].
Bensidhoum, M ;
Chapel, A ;
Francois, S ;
Demarquay, C ;
Mazurier, C ;
Fouillard, L ;
Bouchet, S ;
Bertho, JM ;
Gourmelon, P ;
Aigueperse, J ;
Charbord, P ;
Gorin, NC ;
Thierry, D ;
Lopez, M .
BLOOD, 2004, 103 (09) :3313-3319
[8]   Issue inhibitor of metalloproteinases-3 inhibits shedding of L-selectin from leukocytes [J].
Borland, G ;
Murphy, G ;
Ager, A .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (05) :2810-2815
[9]   Bone marrow endothelial cells increase the invasiveness of human multiple myeloma cells through upregulation of MMP-9: evidence for a role of hepatocyte growth factor [J].
Broek, IV ;
Asosingh, K ;
Allegaert, V ;
Leleu, X ;
Facon, T ;
Vanderkerken, K ;
Van Camp, B ;
Van Riet, I .
LEUKEMIA, 2004, 18 (05) :976-982
[10]   Lymphocyte homing and homeostasis [J].
Butcher, EC ;
Picker, LJ .
SCIENCE, 1996, 272 (5258) :60-66