Three-dimensional cardiac tissue engineering using a thermoresponsive artificial extracellular matrix.
The purpose of this study was to try to reconstitute three-dimensional cardiac tissue using a thermoresponsive artificial extracellular matrix, poly (N-isopropylacrylamide)-grafted gelatin (PNIPAM-gelatin), as the scaffold. PNIPAM-gelatin solution gels almost immediately when heated above 34 degrees C. We thought this property could become advantageous as scaffolding for reconstituting three-dimensional tissue. Because PNIPAM-gelatin solution gels so quickly, all seeded cells in PNIPAM-gelatin solution would become entrapped and uniformly distributed toward three dimensions. Thus it would be possible to reconstitute three-dimensional tissue by a very simple method of mixing cells and PNIPAM-gelatin solution. Fetal rat cardiac cells were mixed with PNIPAM-gelatin solution, incubated at 37 degrees C to allow the mixture to gel, and cultured in vitro. To define suitable culture conditions the following parameters were tested: (1) PNIPAM-gelatin concentration, 0.04 approximately 0.125 mg/ml; (2) cell seeding density, 1 approximately 50 x 10(6) cells/ml; and (3) addition or not of hyaluronic acid. With a PNIPAM-gelatin concentration of 0.05 mg/ml, a cell seeding density of 50 x 10(6) cells/ml, and the addition of hyaluronic acid, tissue was reconstituted and it contracted synchronously. After hematoxylin and eosin staining, the cells reconstituted three-dimensional tissue, and the tissue cross-section was approximately 60 microm thick.
Naito
Hiroshi
H
Department of Artificial Organs, National Cardiovascular Center Research Institute, Fujishiro-dai Suita-city, Osaka, Japan.
Takewa
Yoshiaki
Y
Mizuno
Toshihide
T
Ohya
Shoji
S
Nakayama
Yasuhide
Y
Tatsumi
Eisuke
E
Kitamura
Soichiro
S
Takano
Hisateru
H
Taniguchi
Shigeki
S
Taenaka
Yoshiyuki
Y
D000180:Acrylic Resins / Q000494:pharmacology*; D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D004338:Drug Combinations; D005109:Extracellular Matrix; D005318:Fetal Heart / Q000187:drug effects*; D005333:Fetus; D005780:Gelatin / Q000494:pharmacology*; D006358:Hot Temperature; D006820:Hyaluronic Acid / Q000494:pharmacology; D008858:Microscopy, Phase-Contrast; D009200:Myocardial Contraction; D009206:Myocardium / Q000166:cytology*; D009994:Osmolar Concentration; D051381:Rats; D017208:Rats, Wistar; D012996:Solutions; D013696:Temperature; D023822:Tissue Engineering
D000180:Acrylic Resins; D004338:Drug Combinations; D012996:Solutions; C052970:poly-N-isopropylacrylamide; D005780:Gelatin; D006820:Hyaluronic Acid