Cardiac Differentiation Using BMP4 and CHIR99021
Cardiovascular diseases are the leading cause of death worldwide with approximately 600,000 deaths in the U.S. only. One of the major difficulties with heart diseases is the heart’s failure to produce new and healthy cells in a timely manner. Unfortunately, the heart lacks the ability to quickly generate new tissue composed of viable cells, leading to patient death. Cardiovascular progenitor cells (CVPCs) are believed to be highly useful in cell-dependent therapy aimed at heart diseases. CVPCs are quite similar to stem cells in that they will eventually differentiate into a particular kind of cell, but unlike stem cells, CVPCs are specific and committed to a cell line. Currently, the problem we face with CVPCs is the proper generation and strong development of these cells. Since CVPCs can be derived from human pluripotent stem cells (hPSCs), human induced pluripotent stem cells (hiPSCs), and human embryonic stem cells (hESCs), we attempt to modulate the pathways of converting hPSCs, hiPSCs, and hESCs into CVPCs, thus speeding up the process. We hypothesize that by combining bone morphogenetic protein 4 (BMP4), glycogen synthase kinase 3 (GSK3), inhibitor CHIR99021, and ascorbic acid, we will be able to transform hPSCs, hiPSCs, and hESCs into CVPCs in a much more efficient and timely …show more content…
We freeze the cells when we want to preserve the cells to use at a later date. The steps for freezing cells are: 1) repeat the aforementioned steps 1-9 for passaging stem cells, 2) break up the pellet and place it in 1 or 2 ml of Freezing Medium that has already been cooled down to 4°C, 3) Place in a cryogenic well that has also been cooled to 4°C, 5) Put the vial in isopropanol freezing containers that are initially at room temperature, and 6) place everything in a -80°C freezer for 12-24 hours (Bongso, pg.
Cardiovascular diseases are the leading cause of death worldwide with approximately 600,000 deaths in the U.S. only. One of the major difficulties with heart diseases is the heart’s failure to produce new and healthy cells in a timely manner. Unfortunately, the heart lacks the ability to quickly generate new tissue composed of viable cells, leading to patient death. Cardiovascular progenitor cells (CVPCs) are believed to be highly useful in cell-dependent therapy aimed at heart diseases. CVPCs are quite similar to stem cells in that they will eventually differentiate into a particular kind of cell, but unlike stem cells, CVPCs are specific and committed to a cell line. Currently, the problem we face with CVPCs is the proper generation and strong development of these cells. Since CVPCs can be derived from human pluripotent stem cells (hPSCs), human induced pluripotent stem cells (hiPSCs), and human embryonic stem cells (hESCs), we attempt to modulate the pathways of converting hPSCs, hiPSCs, and hESCs into CVPCs, thus speeding up the process. We hypothesize that by combining bone morphogenetic protein 4 (BMP4), glycogen synthase kinase 3 (GSK3), inhibitor CHIR99021, and ascorbic acid, we will be able to transform hPSCs, hiPSCs, and hESCs into CVPCs in a much more efficient and timely …show more content…
We freeze the cells when we want to preserve the cells to use at a later date. The steps for freezing cells are: 1) repeat the aforementioned steps 1-9 for passaging stem cells, 2) break up the pellet and place it in 1 or 2 ml of Freezing Medium that has already been cooled down to 4°C, 3) Place in a cryogenic well that has also been cooled to 4°C, 5) Put the vial in isopropanol freezing containers that are initially at room temperature, and 6) place everything in a -80°C freezer for 12-24 hours (Bongso, pg.