Project description:A human blastoid, which is an artificial human blastocyst, and has a potential as great tool to investigate fundamentals during the development and establish in vitro models to study the pregnancy failure and birth deficiency, without the use of a human embryo. In 2021, although some methods to generate blastoids have been reported, they used human naïve pluripotent stem cells, which often show genomic instability during in vitro culture. Here, we introduce the simple, robust and scalable method to generate human blastoids from more stable human primed embryonic stem cells. By using non-cell-adhesive hydrogel, hESC aggregates received the chemophysical cellular environment, and forming the asymmetric blastoid structure with the cellular distribution like a human blastocyst. The obtained blastoids also showed the capability the implantation in vitro. This model will enable to elucidate the underlying mechanisms of human pre- and post-implantation process, leading to assisted reproductive technology.
Project description:A human blastoid, which is an artificial human blastocyst, and has a potential as great tool to investigate fundamentals during the development and establish in vitro models to study the pregnancy failure and birth deficiency, without the use of a human embryo. In 2021, although some methods to generate blastoids have been reported, they used human naïve pluripotent stem cells, which often show genomic instability during in vitro culture. Here, we introduce the simple, robust and scalable method to generate human blastoids from more stable human primed embryonic stem cells. By using non-cell-adhesive hydrogel, hESC aggregates received the chemophysical cellular environment, and forming the asymmetric blastoid structure with the cellular distribution like a human blastocyst. The obtained blastoids also showed the capability the implantation in vitro. This model will enable to elucidate the underlying mechanisms of human pre- and post-implantation process, leading to assisted reproductive technology.
Project description:A human blastoid, which is an artificial human blastocyst, and has a potential as great tool to investigate fundamentals during the development and establish in vitro models to study the pregnancy failure and birth deficiency, without the use of a human embryo. In 2021, although some methods to generate blastoids have been reported, they used human naïve pluripotent stem cells, which often show genomic instability during in vitro culture. Here, we introduce the simple, robust and scalable method to generate human blastoids from more stable human primed pluripotent stem cells. By using non-cell-adhesive hydrogel, hPSC aggregates received the chemophysical cellular environment, and forming the asymmetric blastoid structure with the cellular distribution like a human blastocyst. The obtained blastoids also showed the capability the implantation in vitro. This model will enable to elucidate the underlying mechanisms of human pre- and post-implantation process, leading to assisted reproductive technology.
Project description:Mantle cell lymphoma (MCL) is a B cell malignancy characterized by a monoclonal proliferation of lymphocytes with co-expression of CD5, CD43 but not CD23. Typical MCL are associated with cyclin D1 overexpression, and blastoid MCL variants are associated with c-myc translocations. We have developed a murine model of MCL-like lymphoma by crossing Cdk4R24C mice with c-myc-3’RR transgenic mice. Cdk4R24C mice is a knock-in strain that express a Cdk4 protein resistant to inhibition by p16INK4a and other INK4 family members. Breeding Cdk4R24C mice with c-myc-3’RR transgenic mice prone to develop aggressive Burkitt lymphoma-like lymphoma leads in c-myc/Cdk4R24C mice to development of clonal blastoid MCL-like lymphoma. A defect of the INK4-Cdk4 checkpoint can participate to lymphomagenesis in conjunction with additional alterations of cell cycle control, a situation which might be reminiscent of the development of human blastoid MCL. B splenocytes from 4 c-myc/Cdk4(R24C) lymphoma mice and 4 wt mice were investigated.
Project description:human naïve pluripotent stem cells and extended potential stem cells were both used to form blastoid models. We used scRNAseq to investigate the biases founding cell lines have on the formed models.
Project description:Mantle cell lymphoma (MCL) is a B cell malignancy characterized by a monoclonal proliferation of lymphocytes with co-expression of CD5, CD43 but not CD23. Typical MCL are associated with cyclin D1 overexpression, and blastoid MCL variants are associated with c-myc translocations. We have developed a murine model of MCL-like lymphoma by crossing Cdk4R24C mice with c-myc-3’RR transgenic mice. Cdk4R24C mice is a knock-in strain that express a Cdk4 protein resistant to inhibition by p16INK4a and other INK4 family members. Breeding Cdk4R24C mice with c-myc-3’RR transgenic mice prone to develop aggressive Burkitt lymphoma-like lymphoma leads in c-myc/Cdk4R24C mice to development of clonal blastoid MCL-like lymphoma. A defect of the INK4-Cdk4 checkpoint can participate to lymphomagenesis in conjunction with additional alterations of cell cycle control, a situation which might be reminiscent of the development of human blastoid MCL.
Project description:Bulk RNA-seq were performed on E14 EPSCs knockdown during blastoid formation with siRNA, as well as E14 ESC and EPSC treated with T0901317