generation-induced-pluripotent-stem-cells-from-turner-syndrome-45xo

Generation of Induced Pluripotent Stem Cells from Turner Syndrome (45XO) Fetal Cells for Downstream Modelling of Neurological Deficits Associated with the Syndrome

Chromosomal aneuploidies cause severe congenital malformations including central nervous system malformations and fetal death. Prenatal genetic screening ...

Manipal Academy of Higher Education, Manipal, Karnataka, India-576104

A major cause of spontaneous abortions is chromosomal abnormality of foetal cells. We report the generation of an induced pluripotent stem cell line from the fibroblasts isolated from chorionic villi of an early spontaneously aborted foetus with Turner syndrome. The Turner syndrome villus induced pluripotent stem cell line is transgene free, retains the original XO karyotype, expresses pluripotency markers and undergoes trilineage differentiation. This pluripotent stem cell model of Turner syndrome should serve as a tool to study the developmental abnormalities of foetus and placenta that lead to early embryo lethality and profound symptoms like infertility in 45 XO survivors.

Generation of an induced pluripotent stem cell line from chorionic villi of a Turner syndrome spontaneous abortion.

Human term placenta is a bulky organ which harbours abundant mesenchymal stromal cells. This study reports isolation and characterization of placental mesenchymal stromal cells (PMSCs) followed by reprogramming of PMSCs to induced pluripotent stem cells (iPSCs). The placental human iPSC (PhiPSC) line is pluripotent with high telomerase activity, genetically identical to parental PMSCs, transgene free, karyotypically normal and differentiates into ectoderm, mesoderm and endoderm both in vitro and in vivo. Thus PMSCs serve as a unique cell type for human perinatal iPSC derivation from extra embryonic tissue.

Establishment and characterization of induced pluripotent stem cells from placental mesenchymal stromal cells.

Recent approaches of using decellularized organ matrices for cardiac tissue engineering prompted us to culture human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) on the human amniotic membrane (hAM). Since hAM has been used lately to patch diseased hearts in patients and has shown anti-inflammatory and anti-fibrotic benefits, it qualifies as a cardiac compatible and clinically relevant heart tissue scaffold. The aim of this study was to test the ability of the hAM to support attachment, differentiation, and maturation of hiPSC-derived CMs in vitro. hAMs were prepared from term placenta. An in-house generated hiPSC line was used for CM derivation. hiPSC-derived cardiac progenitors were cultured on the surface of cryopreserved hAMs and in the presence of cytokines promoting cardiac differentiation. CMs grown on hAM and popular basement membrane matrix (BMM) Matrigel™ were compared for the following aspects of cardiac development: the morphology of cardiomyocytes with respect to shape and cellular alignments, levels of cardiac-related gene transcript expression, functionality in terms of spontaneous calcium fluxes and mitochondrial densities and distributions. hAM is biocompatible with hiPSC-derived CMs. hAM increased cardiac transcription regulator and myofibril protein transcript expression, accelerated intracellular calcium transients, and enhanced cellular mitochondrial complexity of its cardiomyocytes in comparison to cardiomyocytes differentiated on Matrigel™. Our data suggests that hAM supports differentiation and improves cardiomyogenesis in comparison to Matrigel™. hAMs are natural, easily and largely available. The method of preparing hAM cardiac sheets described here is simple with potential for clinical transplantation. Graphical abstract A An outline of the differentiation protocol with stage-specific growth factors and culture media used. B Cell fates from pluripotent stem cells to cardiomyocytes during differentiation on the amniotic membrane. C-FPhotomicrographs of cells at various stages of differentiation. Scale bars represent 100 μm.

Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis.

Circumcision is a common surgical procedure usually performed on neonates worldwide. Skin removed after circumcision is abundant and often discarded. Neonatal foreskin fibroblasts (NFFs) can be easily isolated from it for application in biomedical research including induced pluripotent stem cell (iPSC) generation. In this study NFFs were reprogrammed to iPSCs using non-integrating episomal reprogramming plasmids. The iPSCs generated express pluripotency markers. They form embryoid bodies and differentiate in vitro into cells of endoderm, mesoderm and ectoderm. They have a normal male karyotype, are genetically identical to the parent fibroblasts and are transgene free.

Article	Year
Generation of an induced pluripotent stem cell line from chorionic villi of a Turner syndrome spontaneous abortion. Stem cell research\| PubMed ID: 28412999	2017
Establishment and characterization of induced pluripotent stem cells from placental mesenchymal stromal cells. Stem cell research\| PubMed ID: 29291511	2018
Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis. In vitro cellular & developmental biology. Animal\| PubMed ID: 30798515	2019
Generation of integration free human iPSC line SORMi002-A from neonatal foreskin fibroblasts. Stem cell research\| PubMed ID: 31454679	2019

Shagufta Parveen

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