A Simplified and Effective Approach for the Isolation of Small Pluripotent Stem Cells Derived from Human Peripheral Blood

Pluripotent stem cells are key players in regenerative medicine. Embryonic pluripotent stem cells, despite their significant advantages, are associated with limitations such as their inadequate availability and the ethical dilemmas in their isolation and clinical use. The discovery of very small embryonic-like (VSEL) stem cells addressed the aforementioned limitations, but their isolation technique remains a challenge due to their small cell size and their efficiency in isolation. Here, we report a simplified and effective approach for the isolation of small pluripotent stem cells derived from human peripheral blood. Our approach results in a high yield of small blood stem cell (SBSC) population, which expresses pluripotent embryonic markers (e.g., Nanog, SSEA-3) and the Yamanaka factors. Further, a fraction of SBSCs also co-express hematopoietic markers (e.g., CD45 and CD90) and/or mesenchymal markers (e.g., CD29, CD105 and PTH1R), suggesting a mixed stem cell population. Finally, quantitative proteomic profiling reveals that SBSCs contain various stem cell markers (CD9, ITGA6, MAPK1, MTHFD1, STAT3, HSPB1, HSPA4), and Transcription reg complex factors (e.g., STAT5B, PDLIM1, ANXA2, ATF6, CAMK1). In conclusion, we present a novel, simplified and effective isolating process that yields an abundant population of small-sized cells with characteristics of pluripotency from human peripheral blood.

Automated summary

Pluripotent stem cells play a vital role in regenerative medicine. However, the limited availability and ethical concerns associated with embryonic pluripotent stem cells have led to the discovery of very small embryonic-like (VSEL) stem cells. In this study, a simplified and effective method for isolating small pluripotent stem cells from human peripheral blood is presented. The isolated small blood stem cells (SBSC) population expresses pluripotency markers and also exhibits characteristics of hematopoietic and mesenchymal stem cells. Proteomic profiling reveals the presence of various stem cell markers and transcription regulatory complex factors in SBSCs. This novel isolation process yields a abundant population of small-sized cells with pluripotent characteristics from human peripheral blood.