Altered migration and adhesion potential of pro-neurally converted human bone marrow stromal cells.
BACKGROUND
Bone marrow (BM)-derived mesenchymal stromal cells (MSC) are promising candidate cells for the development of neuroregenerative therapies. We have previously introduced the pro-neural conversion of human MSC to neural stem cell-like cells (m-NSC) by culturing them in suspension culture under serum-free conditions.


METHODS
In the present study, we used a modified Boyden chamber assay to study the influence of various chemoattractants and extracellular matrix components on MSC and m-NSC migration in vitro. The underlying mechanisms were investigated further by applying real-time reverse transcriptase (RT)-polymerase chain reaction (PCR) and flow cytometry.


RESULTS
The basal migration of m-NSC was significantly reduced compared with MSC (six versus 27 out of 10,000 cells migrated within 6 h). We evaluated the effects of bone morphogenic protein 2 (BMP2), insulin-like growth factor 1 (IGF1), platelet-derived growth factor bb (PDGFbb), vascular endothelial growth factor (VEGFa), and stromal cell-derived factor 1 (SDF1) on the migration potential of both cell types and PDGFbb proved to be the most potent stimulant of migration (235 versus 198 m-NSC or MSC migrated). Adhesion of m-NSC to the filter membrane was delayed and not affected by IGF1 or PDGFbb: 90% of MSC, but only 20% of m-NSC, adhered within 1 h, with 90% of m-NSC adhering within 3 h. However, real-time RT-PCR and flow cytometry revealed an up-regulation of the PDGF receptor B following conversion. Coating the membranes with collagen type I or hyaluronan also significantly influenced cell migration.


CONCLUSIONS
We could identify major chemoattractive factors for m-NSC and gained partial insight into the complex processes involved in migration of neurally converted cells.
Habisch
H-J
HJ
Department of Neurology, University of Ulm, Ulm, Germany.
Fiedler
J
J
Ludolph
A C
AC
Storch
A
A
Brenner
R E
RE
D000293:Adolescent; D000328:Adult; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002448:Cell Adhesion / Q000235:genetics; D018929:Cell Culture Techniques; D002465:Cell Movement / Q000235:genetics; D054338:Cell Transdifferentiation / Q000235:genetics; D054377:Chemokine CXCL12 / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008875:Middle Aged; D009420:Nervous System / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D020574:Proto-Oncogene Proteins c-sis / Q000378:metabolism*; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D054377:Chemokine CXCL12; D024042:Collagen Type I; D020574:Proto-Oncogene Proteins c-sis; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D007334:Insulin-Like Growth Factor I; D006820:Hyaluronic Acid; D020797:Receptor, Platelet-Derived Growth Factor beta