Developmentally Conserved CDX2 Cells from Human Term Placenta Drive Cardiovascular Regeneration

We report a population of multipotent cells isolated from term human placentas that exhibit clonal expansion, migratory capacity, and a gene expression profile indicating immune privilege. These cells, expressing the developmental regulator CDX2, previously known only for its role in early placentation, differentiate into cardiomyocytes and vascular cells. Building on our prior findings that murine Cdx2 cells improved cardiac function in mice after myocardial infarction (MI), we isolated CDX2⁺ cells from placentas of 180 healthy pregnancies. These human CDX2 cells spontaneously generate cardiac and vascular lineages in vitro, in vivo, and express transcriptomic signatures associated with cardiogenesis, vasculogenesis, immune modulation, and chemotaxis. When administered to NOD/SCID mice after MI, the cells restore cardiac function. Additionally, CDX2 cells can be clonally propagated while retaining cardiovascular differentiation potential. Our findings support the therapeutic potential of placental CDX2 cells as an ethically accessible and regenerative strategy for cardiovascular disease. Human CDX2mCherry-positive cell samples, both freshly isolated (n=2) and expanded at passage 3 (n=3), were thawed for analysis (total n=5). Cell counts were assessed using the Cellaca Cell Counter (Nexcelom Bioscience, Lawrence, MA, USA) in conjunction with NucBlue for staining cell nuclei and propidium iodide (PI) for identifying dead cells. Samples were then processed with the Chromium X platform (10x Genomics, Pleasanton, CA, USA), following the manufacturer’s protocols, to target the capture of approximately 3,000 cells per sample for downstream analysis. Single-cell RNA-seq libraries were prepared using the Chromium Next GEM Single Cell 3’ Gene Expression Kit. Library quality was assessed using the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified via Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA, USA). Additional quantification was performed using qPCR (Applied Biosystems, Carlsbad, CA, USA) before sequencing on an Illumina platform, adhering to 10x Genomics' recommended sequencing configurations