Project description:global gene expression were compared among human blood iPSC, human fibroblas iPSC, human embryonic stem cells, human bone marrow MNC and human forskin fibroblast Reprogramming blood cells to induced pluripotent stem cells (iPSCs) provides a novel tool for modeling blood diseases in vitro. we demonstrated that iPSCs free of transgene and vector sequences could be efficiently generated from human bone marrow and cord blood mononuclear cells using non-integrating episomal vectors. The reprogramming described here is up to 100 times more efficient, occurs 1 to 3 weeks faster as compared to the reprogramming of fibroblasts, and does not require isolation of progenitors or multiple rounds of transfection. This approach provides an opportunity to explore banked normal and diseased cord blood and bone marrow samples without the limitations associated with virus-based methods. Compare the global gene expression of iPSs from different sources, ESCs and Somatic cells
Project description:global gene expression were compared among human blood iPSC, human fibroblas iPSC, human embryonic stem cells, human bone marrow MNC and human forskin fibroblast Reprogramming blood cells to induced pluripotent stem cells (iPSCs) provides a novel tool for modeling blood diseases in vitro. we demonstrated that iPSCs free of transgene and vector sequences could be efficiently generated from human bone marrow and cord blood mononuclear cells using non-integrating episomal vectors. The reprogramming described here is up to 100 times more efficient, occurs 1 to 3 weeks faster as compared to the reprogramming of fibroblasts, and does not require isolation of progenitors or multiple rounds of transfection. This approach provides an opportunity to explore banked normal and diseased cord blood and bone marrow samples without the limitations associated with virus-based methods.
Project description:Reprogramming of somatic cells provides potential for the generation of specific cell types, which could be a key step in the study and treatment of human diseases. In vitro reprogramming of somatic cells into a pluripotent embryonic stem (ES) cellM-bM-^@M-^Slike state has been reported by retroviral transduction of murine fibroblasts using four embryonic transcription factors or through cell fusion of somatic and pluripotent stem cells. The generation of reprogrammed pluripotent cells using a somatic cell donor source such as bone marrow (BM) or peripheral blood is of particular therapeutic interest because of the relative ease of harvesting these cell types. Here we show that mouse adult BM mononuclear cellsM-oM-<M-^HBM MNCsM-oM-<M-^Iare competent as donor cells and can be reprogrammed into pluripotent ES cell-like cells. We isolated BM MNCs and embryonic fibroblasts (MEFs) from Oct4-EGFP transgenic mice, fused them with ES cells and infected them with retroviruses expressing Oct4, Sox2, Klf4, and c-Myc. Fused BM cells formed more ES-like colonies than did MEFs. Infected BM cells gave rise to iPS cells, although transduction efficiencies were not high. It was more efficient to pick up iPS colonies as compared with MEFs. BM-derived iPS (BM iPS) cells expressed embryonic stem cell markers, formed teratomas, and contributed to chimera mice with germline development. Clonal analysis revealed that BM iPS clones had diversity, although some clones were found to be genetically identical with different phenotypes. Here we demonstrate, for the first time, the induction of pluripotent cells directly from hematopoietic tissue. Gene expression profiling was performed in mouse BMMNCs, ES and BMMNC derived iPS cell lines.
Project description:Induced pluripotent stem (iPS) cells can be generated from somatic cells by transduction with several transcription factors in both mouse and human. However, direct reprogramming in other species has not been reported. Here, we established an efficient method to generate monkey iPS cells from fibroblasts by retrovirus-mediated introduction of the four monkey transcription factors OCT4 (POU5F1), SOX2, KLF4, and c-MYC. The monkey iPS cells displayed ES-like morphology, expressed ES cell-marker genes, shared similar global gene profiles and methylation status in the OCT4 promoter to those of monkey ES cells, and possessed the ability to differentiate into three germ layers in vitro and in vivo. Our results suggest that the mechanism of direct reprogramming is conserved among species. The efficient generation of monkey iPS cells will allow investigation of the feasibility of therapeutic cloning in primate model with various diseases. Keywords: Induced pluripotent stem, iPS, Rhesus monkey We analysed each sample (Rhesus monkey fibroblast, embryonic stem cell (ES) and induced pluripotent stem cell (iPS)) for three replications and sought to see high similarty between iPS and ES.
Project description:Human induced pluripotent stem cells provide an unlimited, scalable source of youthful tissue progenitors and secretome for regenerative therapies. The aim of our study was to assess the potential of conditioned medium (CM) derived from hiPSC-mesenchymal progenitors (hiPSC-MPs) to stimulate osteogenic differentiation of adult and aged human bone marrow-mesenchymal stromal cells (MSCs). In addition, we evaluated whether extended cultivation or osteogenic pre-differentiation of hiPSC-MPs could enhance the CM stimulatory activity.
Project description:Efficient Generation of Transgene-Free Induced Pluripotent Stem Cells from Normal and Neoplastic Bone Marrow and Cord Blood Mononuclear Cells
Project description:Cell fate manipulation is powerful for generating desired cell types through reprogramming, such as induced pluripotent stem cells (iPSCs), with broad biological applications. However, reprogramming can be risky due to dramatic changes in cell identity, necessitating strict regulation to ensure safety and efficacy. p53 is essential for maintaining genome stability to prevent abnormalities induced by changes of cell states, which however, functionally counters Yamanaka factors for efficient reprogramming. Thus, delicately balancing p53 activity for efficient reprogramming with highly quality has proven challenging. Here, we demonstrate that efficient human chemical reprogramming is safeguarded by p53, in distinction from Yamanaka factors-mediated reprogramming. Suppressing p53 activity unexpectedly and substantially hinders the generation of chemically induced pluripotent stem cells (CiPSCs). Notably, p53 precisely regulates the mesenchymalization occurred at the early reprogramming stage where cell plasticity is re-established and curbs excessive epithelial-mesenchymal transition (EMT). Furthermore, the retinoid acid receptor agonist, TTNPB, leverages p53’s anti-metastatic functions to facilitate CiPSC generation by activating BTG2. Our results demonstrate that p53 activity distinguishes chemical reprogramming from Yamanaka factors-mediated reprogramming, and offer a promising strategy that delicately balances p53 with reprogramming for both safe and efficient cell fate manipulation.
Project description:Cell fate manipulation is powerful for generating desired cell types through reprogramming, such as induced pluripotent stem cells (iPSCs), with broad biological applications. However, reprogramming can be risky due to dramatic changes in cell identity, necessitating strict regulation to ensure safety and efficacy. p53 is essential for maintaining genome stability to prevent abnormalities induced by changes of cell states, which however, functionally counters Yamanaka factors for efficient reprogramming. Thus, delicately balancing p53 activity for efficient reprogramming with highly quality has proven challenging. Here, we demonstrate that efficient human chemical reprogramming is safeguarded by p53, in distinction from Yamanaka factors-mediated reprogramming. Suppressing p53 activity unexpectedly and substantially hinders the generation of chemically induced pluripotent stem cells (CiPSCs). Notably, p53 precisely regulates the mesenchymalization occurred at the early reprogramming stage where cell plasticity is re-established and curbs excessive epithelial-mesenchymal transition (EMT). Furthermore, the retinoid acid receptor agonist, TTNPB, leverages p53’s anti-metastatic functions to facilitate CiPSC generation by activating BTG2. Our results demonstrate that p53 activity distinguishes chemical reprogramming from Yamanaka factors-mediated reprogramming, and offer a promising strategy that delicately balances p53 with reprogramming for both safe and efficient cell fate manipulation.
Project description:Cell fate manipulation is powerful for generating desired cell types through reprogramming, such as induced pluripotent stem cells (iPSCs), with broad biological applications. However, reprogramming can be risky due to dramatic changes in cell identity, necessitating strict regulation to ensure safety and efficacy. p53 is essential for maintaining genome stability to prevent abnormalities induced by changes of cell states, which however, functionally counters Yamanaka factors for efficient reprogramming. Thus, delicately balancing p53 activity for efficient reprogramming with highly quality has proven challenging. Here, we demonstrate that efficient human chemical reprogramming is safeguarded by p53, in distinction from Yamanaka factors-mediated reprogramming. Suppressing p53 activity unexpectedly and substantially hinders the generation of chemically induced pluripotent stem cells (CiPSCs). Notably, p53 precisely regulates the mesenchymalization occurred at the early reprogramming stage where cell plasticity is re-established and curbs excessive epithelial-mesenchymal transition (EMT). Furthermore, the retinoid acid receptor agonist, TTNPB, leverages p53’s anti-metastatic functions to facilitate CiPSC generation by activating BTG2. Our results demonstrate that p53 activity distinguishes chemical reprogramming from Yamanaka factors-mediated reprogramming, and offer a promising strategy that delicately balances p53 with reprogramming for both safe and efficient cell fate manipulation.