Isolation and culture of adult and fetal rabbit bladder smooth muscle cells and their interaction with biopolymers.

Department of Pediatric Surgery, Kocaeli University Medical Faculty, Kocaeli, Turkey.
Journal of Pediatric Surgery (Impact Factor: 1.38). 02/2003; 38(1):21-4. DOI:10.1053/jpsu.2003.50003
Source: PubMed

ABSTRACT The aim of this study was to test the feasibility of isolation and culture of adult and fetal rabbit bladder smooth muscle cells (SMCs) and comparison of their interactions with different types of biodegradable biopolymers in cell culture.
Bladder SMCs isolated from adult and fetus rabbits were identified by immunostaining for smooth muscle alpha-actin and myosin. Growth kinetics of SMCs were estimated using population doubling time (PDT) and thymidine labeling index (TLI). Poly (D, L-lactide-co-glycolide; PLGA) copolymers were synthesized at 85:15 and 75:25 monomer ratios. The porous scaffolds prepared from these polymers were seeded with SMCs. The study compared the effectiveness of adsorbing fibronectin and fetal calf serum (FCS) on these biopolymers. The cells grown on these polymers were quantified using a neutral red uptake assay.
Over 90% of the 2 cell populations stained positive for SMC marker proteins. Fetal SMCs were seen to emerge from the tissue after 3 to 4 days, whereas adult SMCs were seen after 5 to 6 days. However, estimated PDT of fetal and adult SMCs was 85.2 and 54.5 hours, respectively, and TLI of adult SMCs was also higher than with fetal SMCs. Proliferation on 75:25 PLGA was better than for 85:15 and for both biopolymers; adsorption of FCS significantly affected cell attachment relative to fibronectin.
Although fetal SMCs were shown to emerge from explants early after seeding onto dishes, doubling time and S-phase fraction of adult bladder SMCs were markedly higher than of fetal derived cells. Adsorption of serum proteins significantly enhances the attachment of both fetal and adult SMCs to biopolymers.

0 0
  • [show abstract] [hide abstract]
    ABSTRACT: The application of stem cells and their use in tissue-engineering approaches is emerging in clinical therapeutic intervention strategies. The use of adult stem cells, either autologous or allogenic, does not raise ethical concerns, in contrast to embryonic stem cells. Mesenchymal stromal cells (MSCs) can be easily obtained from bone marrow or from adipose tissue and further expanded in vitro. Due to their differentiation capacity, MSCs are very attractive for tissue engineering purposes. Furthermore, MSCs secrete a variety of mediators that have beneficial effects on the regenerating tissue. In this review we give an insight into stem cell hierarchy, define the properties of MSCs and summarize recent reports of their administration in urological diseases.
    BJU International 11/2009; 105(3):309-12. · 3.05 Impact Factor
  • Source
    [show abstract] [hide abstract]
    ABSTRACT: A PLGA biodegradable membrane can be used as a scaffold for mucociliary epithelium transfer. The aim of this study was to examine the usefulness of the PLGA membrane as a biodegradable scaffold for mucociliary epithelium transfer in order for it to be used as a substitute for a skin graft for restoring mucosal defects in the airway. A PLGA biodegradable membrane was synthesized using the immersion precipitation method, and morphologic characterization was carried out using scanning electron microscopy (SEM). The degradation test was performed by soaking the PLGA membrane in a culture medium, and the morphological changes were observed by SEM. Human nasal basal epithelial (HNBE) cells were cultured on the newly synthesized PLGA membrane, and the morphological changes were analyzed using SEM. The MUC5AC and MUC8 mRNA levels were analyzed by RT-PCR. The PLGA membrane for the mucociliary epithelium transfer was successfully fabricated. It had a 24 mm diameter, a 50 microm thickness, and many pores with diameters of approximately 3 microm. The PLGA membrane began to degrade from 7 days after it was soaked in the culture medium. It rapidly degraded from 3 weeks and severe destruction of the pore structure was noted from 4 to 6 weeks of soaking. The HNBE cells were well differentiated into the mucociliary epithelium on the PLGA membrane both phenotypically and genotypically.
    Acta Oto-Laryngologica 07/2006; 126(6):594-9. · 1.11 Impact Factor


Available from
Sep 13, 2013