Effect of poly-L-lysine coating on retinoic acid-loaded PLGA microspheres in the differentiation of carcinoma stem cells into neural cells.
In this study, PLGA microspheres were prepared using a water-in-oil-in-water emulsion/solvent evaporation technique. Some microspheres were coated with poly-L-lysine (an extracellular matrix (ECM) component), and then pluripotent P19 embryonic carcinoma cells were seeded on them. P19 cells attached onto the PLGA microspheres; subsequently, by adding retinoic acid (RA) to cell culture medium as a neurogenic inducer (RA was released from the microspheres), the cells differentiated into neural cells. Size and morphology of PLGA microspheres was characterized by scanning electron microscopy (SEM). Neurogenic differentiation was studied by immunofluorescent staining, real-time polymerase chain reaction (RT-PCR), and light microscopy. Histological assay showed that more cells attached onto microspheres coated with poly-L-lysine than the uncoated group. Immunofluoresent staining and RT-PCR analysis for ß-Tubulin, Nestin and Pax6 genes indicated differentiation of P19 cells into neural cells on both coated and uncoated microspheres. It was found that a high surface area of microspheres improves cell attachment and expansion, which was significantly increased in those coated with poly-L-lysine. Finally, these results highlight the versatility of these sample scaffolds as a model system for nerve tissue engineering.
Stem Cell Department, Cell Science Research Center, Royan Institute, Tehran, Iran.
Hanieh
H
Nojehdehian
Fathollah
F
Moztarzadeh
Hossein
H
Baharvand
Narges Zare
NZ
Mehrjerdi
Hamid
H
Nazarian
Mohammadreza
M
Tahriri
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D004339:Drug Compounding; D005136:Eye Proteins / Q000235:genetics; D005455:Fluorescent Antibody Technique; D018398:Homeodomain Proteins / Q000235:genetics; D007381:Intermediate Filament Proteins / Q000235:genetics; D007700:Kinetics; D019344:Lactic Acid / Q000737:chemistry*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009419:Nerve Tissue Proteins / Q000235:genetics; D064231:Nestin; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics; D010316:Particle Size; D039904:Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011107:Polylysine / Q000737:chemistry*; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012995:Solubility; D013499:Surface Properties; D013997:Time Factors; D054457:Tissue Scaffolds; D014212:Tretinoin / Q000737:chemistry / Q000494:pharmacology*; D014404:Tubulin / Q000235:genetics
D005136:Eye Proteins; D018398:Homeodomain Proteins; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D012097:Repressor Proteins; D014404:Tubulin; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011107:Polylysine; D011100:Polyglycolic Acid; D019344:Lactic Acid; D014212:Tretinoin