Differentiation of adult stem cells into smooth muscle for vascular tissue engineering.
BACKGROUND
Herein we evaluate the potential of adipose-derived stem cells (ASC) to differentiate into smooth muscle cells (SMC) and their potential for use in a tissue-engineered vascular graft.


METHODS
We isolated ASC (CD13+29+90+) from the peri-umbilical adipose tissue of patients undergoing vascular surgery, and cultured them in media containing angiotensin II (AngII), sphingosylphosphorylcholine (SPC), or transforming growth factor-beta 1 (TGFβ1) for up to 3 weeks. SMC differentiation was assessed by (1) expression of early (calponin, caldesmon) and late (myosin heavy chain, MHC) SMC markers by RT-PCR, qPCR and Western blot, and (2) contraction upon plating on collagen gel. Differentiated ASCs were seeded onto a vascular graft (decellularized saphenous vein) within a bioreactor, and cell attachment was determined using confocal microscopy.


RESULTS
Prior to differentiation, ASC expressed low levels of all three molecular markers. After culture in each differentiating medium, the extent of up-regulation of calponin, caldesmon, and MHC was variable across all cell lines. After seeding onto collagen gel, ASCs differentiated in SPC and TGFβ1 exhibit contractile properties, similar to smooth muscle cell controls. Differentiated stem cells adhered and proliferated on the vascular graft.


CONCLUSIONS
These data suggest that human adipose-derived stem cells (1) exhibit variable expression of SMC molecular markers after differentiation, (2) exhibit a contractile phenotype after differentiation with SPC and TGFβ1, and (3) proliferate on a vascular graft scaffold. Thus, ASCs are potentially useful in the construction of autologous arteries.
Harris
Lisa J
LJ
Department of Surgery, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Abdollahi
Hamid
H
Zhang
Ping
P
McIlhenny
Stephen
S
Tulenko
Thomas N
TN
DiMuzio
Paul J
PJ
D053687:Adult Stem Cells / Q000187:drug effects*; D000368:Aged; D000369:Aged, 80 and over; D000804:Angiotensin II / Q000494:pharmacology*; D001807:Blood Vessel Prosthesis; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D003094:Collagen; D005260:Female; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D010641:Phenotype; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D000096985:Calponins
D000804:Angiotensin II; D002135:Calcium-Binding Proteins; D002148:Calmodulin-Binding Proteins; D003094:Collagen; D008840:Microfilament Proteins; D018995:Myosin Heavy Chains; D010767:Phosphorylcholine; D013110:Sphingosine; D016212:Transforming Growth Factor beta; D000096985:Calponins; C005356:sphingosine phosphorylcholine