TY - JOUR
T1 - Co-culture of human cardiomyocyte and human amnion epithelial stem cells in amnion bilayer matrix for cardiomyogenesis
AU - Putra, Muhammad A.
AU - Sandora, Normalina
AU - Kusuma, Tyas R.
AU - Fitria, Nur A.
AU - Soetisna, Tri W.
AU - Busro, Pribadi W.
AU - Ardiansyah,
AU - Muttaqin, Chaidar
AU - Makdinata, William
AU - Alwi, Idrus
N1 - Funding Information:
This study was supported by grant no. NKB-0541/UN2.R3.1/ HKP.05.00/2019 from the University of Indonesia.
Publisher Copyright:
© 2022, Universitatsklinikum Hamburg - Eppendorf. All rights reserved.
PY - 2022/6/30
Y1 - 2022/6/30
N2 - Our study developed a culture system to deliver cells for the regeneration of infarcted myocardial tissue. Human cardiomyocytes (hCardio) and human amnion epithelial stem cells (hAESC) were co-cultured in a biologi-cal scaffold forming a 3-D matrix, prepared as a heart patch candidate. The current investigation was aimed for assessment of hCardio-to-hAESC seeding ratio for the best conditions to provide cardiomyogenesis, i.e., 1:5 or the 1:6. This ratio corresponded to optimal cell number of 500,000 cells/cm2, thus providing a 12-cm2 heart patch. This cell ratio was also published in our previous study using human adipose stem cells and hCardio. The hCardio was isolated from the heart tissue taken during surgical correction performed in patients with right ventricular hypertrophy. The cell isolates expressed cTnT (10.7%), cKit/CD117 (16%), ICAM (94%), and PECAM+/VCAM-33%. The hAESC was isolated from an elected donor at Caesarean section. It showed expression of TRA-1-60 (82.4%), SSEA-4 (28.2%), Oct-3/4 (2.9%), Nanog (11.4%), without expression of immune antigens, e.g., HLA-DR (0%), HLA-ABC (0.2%) as well as low-level expression of mesenchymal stem cell (MSC) markers, i.e., CD73 (20.2%), CD90 (0.4%), CD105 (59.2%). These cells did not exhibit Lin phenotype (CD3, CD14, CD16, CD19, CD20, CD56). At both seeding ratios, the co-culture matrix also re-leased TNF-α at low levels (<1 pg/ml) throughout cultivation from day 1 to day 8. Both groups had shown a consistent growth of cells over time. Confocal images showed that the cell population has expanded and mi-grated to the deeper plane up to 140 µm after 5 days of incubation in both 1:5 and 1:6 groups. The cells seemed to connect and form projections, starting by day 5, being more obvious among the 1:5 group. They proliferated rapidly until no solitary cells were observed after day 8 of co-culture. Expression of the cardiomyogenesis genes, e.g., cTnT and ACTN2 in the 1:6 group were expressed, similarly to normal cardiomyocytes after eight days of cultivation. MHC genes in the 1:6 group were also ex-pressed, although less than 1-fold to normal cardiomy-ocytes. This study indicated that hCardio and hAESC at 1:6 ratio seeded on amnion bilayer could support cardi-omyogenesis derivation from the progenitor cells.
AB - Our study developed a culture system to deliver cells for the regeneration of infarcted myocardial tissue. Human cardiomyocytes (hCardio) and human amnion epithelial stem cells (hAESC) were co-cultured in a biologi-cal scaffold forming a 3-D matrix, prepared as a heart patch candidate. The current investigation was aimed for assessment of hCardio-to-hAESC seeding ratio for the best conditions to provide cardiomyogenesis, i.e., 1:5 or the 1:6. This ratio corresponded to optimal cell number of 500,000 cells/cm2, thus providing a 12-cm2 heart patch. This cell ratio was also published in our previous study using human adipose stem cells and hCardio. The hCardio was isolated from the heart tissue taken during surgical correction performed in patients with right ventricular hypertrophy. The cell isolates expressed cTnT (10.7%), cKit/CD117 (16%), ICAM (94%), and PECAM+/VCAM-33%. The hAESC was isolated from an elected donor at Caesarean section. It showed expression of TRA-1-60 (82.4%), SSEA-4 (28.2%), Oct-3/4 (2.9%), Nanog (11.4%), without expression of immune antigens, e.g., HLA-DR (0%), HLA-ABC (0.2%) as well as low-level expression of mesenchymal stem cell (MSC) markers, i.e., CD73 (20.2%), CD90 (0.4%), CD105 (59.2%). These cells did not exhibit Lin phenotype (CD3, CD14, CD16, CD19, CD20, CD56). At both seeding ratios, the co-culture matrix also re-leased TNF-α at low levels (<1 pg/ml) throughout cultivation from day 1 to day 8. Both groups had shown a consistent growth of cells over time. Confocal images showed that the cell population has expanded and mi-grated to the deeper plane up to 140 µm after 5 days of incubation in both 1:5 and 1:6 groups. The cells seemed to connect and form projections, starting by day 5, being more obvious among the 1:5 group. They proliferated rapidly until no solitary cells were observed after day 8 of co-culture. Expression of the cardiomyogenesis genes, e.g., cTnT and ACTN2 in the 1:6 group were expressed, similarly to normal cardiomyocytes after eight days of cultivation. MHC genes in the 1:6 group were also ex-pressed, although less than 1-fold to normal cardiomy-ocytes. This study indicated that hCardio and hAESC at 1:6 ratio seeded on amnion bilayer could support cardi-omyogenesis derivation from the progenitor cells.
KW - 3-D scaffold
KW - amnion bilayer
KW - cardiomyogenesis
KW - cell therapy
KW - Human amnion epithelial stem cells
KW - human cardiomy-ocytes
UR - http://www.scopus.com/inward/record.url?scp=85135755698&partnerID=8YFLogxK
U2 - 10.18620/ctt-1866-8836-2022-11-2-72-83
DO - 10.18620/ctt-1866-8836-2022-11-2-72-83
M3 - Article
AN - SCOPUS:85135755698
SN - 1867-416X
VL - 11
SP - 72
EP - 83
JO - Cellular Therapy and Transplantation
JF - Cellular Therapy and Transplantation
IS - 2
ER -
