Protocol For Differentiating Pluripotent Stem Cells Into Primordial Germ Cell-like Cells.

Protocol of Human Primordial Germ Cell-like Cell Generation from Pluripotent Stem Cells in 2D and 3D Culture Systems

Simple Summary Human in vitro gametogenesis is the focus of researchers, as pluripotent stem cell derived human primordial germ cell-like cells (hPGCLCs) could not complete the meiotic division. Therefore, generating early hPGCLCs, which provides an opportunity for further investigations to overcome a meiotic block, is a cue of success in deriving haploid gametes in vitro. Thus, we described a protocol for hPGCLC specification of human pluripotent stem cells (hPSCs) through sequential induction with Activin A for 2 days and BMP4 for 6 days in 2D and 3D culture systems. Induction of hPSCs into hPGCLCs demonstrated expression of early primordial germ cell markers, including PRDM1, NANOS3, DAZL, STELLA, SOX17, SSEA1, and cKIT, on the 8th day of hPGCLC generation.

Harnessing Endogenous Plasticity Rather than Reprogramming of Mature Cells Will Advance Regenerative Medicine, Cancer Treatment and Rejuvenation

Recognising the central role of VSELs/progenitors and their niche in maintaining tissue homeostasis in vivo could resolve existing roadblocks and guide more effective endogenous regenerative therapies for diseased tissues and age-related dysfunctions.

Characterization of the metabolome and proteome of stem cell-derived human primordial germ cell-like cells: a multi-omics approach.

Primordial germ cells (PGCs) are the population of cells that, in the human embryo, are initially specified at day 12 post-fertilization, and form the precursor cells for the future gametes. Although in vitro differentiation of PGCs from human stem cells has been achieved, these primordial germ cell-like cells (hPGCLCs) fail to completely mature without the use of ex vivo human or animal gonadal soma. Previous studies in mice revealed that several metabolic changes occur during the specification and maturation of these cells, which are essential for their developmental progress. However, little is known about the metabolic profile of human primordial germ cells. In the scarcity of human PGCs, particularly at the early specification stage, hPGCLCs serve as a research model to study PGC formation. To characterize the metabolic and proteomic profile of these cells, we differentiated hPGCLCs using induced-pluripotent stem cells and performed a mass spectrometry analysis to establish their metabolome and proteome. These cells revealed distinct metabolic profile, with changes particularly at the proteome level. This included a shift between canonical and non-canonical citric acid cycle in hPGCLCs, downregulation of late-stage glycolysis and reduction of nucleotide de novo synthesis. By providing an integrative map of these metabolic networks, we aim to provide insight on the metabolism of hPGCLC development that could help improve methods for fully in vitro differentiation and maturation of hPGCLCs.

Advances in Induced Pluripotent Stem Cell

Because they can propagate indefinitely, as well as give rise to every other cell type in the body (such as neurons, heart, pancreatic, and liver cells), they represent a single source of cells that could be used to replace those lost to damage or disease.

Rapid and efficient generation of human 8-cell-like cells for embryo modelling

Modeling myeloid cell development in health and disease using induced pluripotent stem cells

Key applications of iPSC technology in hemato-oncology are summarized, its major advantages and current limitations are discussed, and emerging directions are highlighted, including scalable iPSC-derived blood cell therapies for inherited and acquired bone marrow failure syndromes and leukemia.