title: Genetically modified adipose tissue-derived mesenchymal stem cells overexpressing CXCR4 display increased motility, invasiveness, and homing to bone marrow of NOD/SCID mice.
abstract: OBJECTIVE
  This study evaluates usefulness of CXCR4 overexpression via retroviral transduction in adipose tissue-derived mesenchymal stem cells (AT-MSCs) as a strategy to increase their migration and engraftment ability.
  
  
  METHODS
  AT-MSCs were isolated from lipoaspirates from human healthy donors with liberase 3. Cells were transduced with retroviral vector carrying either CXCR4 or green fluorescent protein (GFP) complementary DNA, and neo-resistant colonies were selected and used in experiments. Chemotaxis, invasion through Matrigel, motor activity, gene expression, osteodifferentiation potential, and engraftment into bone marrow of nonobese diabetic/severe combined immunodeficient mice were analyzed for CXCR4-overexpressing cells and GFP-control cells.
  
  
  RESULTS
  Approximately 90% of retrovirus-transduced AT-MSCs expressed CXCR4 or GFP and maintained their ability to differentiate into osteocytes. CXCR4-transduced AT-MSCs displayed enhanced migration and higher invasiveness toward SDF-1 gradient. The upregulation of CXCR4 led to phosphorylation of mitogen-activated protein and AKT kinases and an increase in metalloproteinase expression after SDF-1 stimulation. The transplantation of CXCR4-transduced AT-MSCs into nonobese diabetic/severe combined immunodeficient mice led to increased engraftment into bone marrow in comparison to GFP-transduced AT-MSCs.
  
  
  CONCLUSIONS
  Adipose tissue is one of the alternative sources of MSCs to bone marrow. We showed that AT-MSCs overexpressing CXCR4 preserve their ability for osteodifferentiation. Enhanced migration and engraftment of the transduced AT-MSCs into bone marrow indicate the usefulness of this strategy in overcoming low engraftment of MSCs in clinical approaches of cellular therapies for bone disorders and can represent a powerful tool in regenerative medicine and gene therapies. Thus, these cells may be used as an alternative to bone marrow-derived MSCs.
authors.affiliation: Department of Transplantation, Collegium Medicum, Jagiellonian University, Cracow, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Bobis-Wozowicz
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Miekus
authors.forename: Ewa
authors.initials: E
authors.lastname: Wybieralska
authors.forename: Danuta
authors.initials: D
authors.lastname: Jarocha
authors.forename: Artur
authors.initials: A
authors.lastname: Zawisz
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Madeja
authors.forename: Marcin
authors.initials: M
authors.lastname: Majka
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002465:Cell Movement / Q000235:genetics*; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008875:Middle Aged; D019718:Receptors, CXCR4 / Q000235:genetics*; D015398:Signal Transduction
chemical_list: C514936:CXCR4 protein, mouse; D019718:Receptors, CXCR4