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Stem cell challenges and opportunities.

Kenta Ite1, Masashi Toyoda2, Saeko Akiyama3

  • 1Center for Regenerative Medicine, National Center for Child Health and Development Research Institute, Tokyo, Japan; Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.

Progress in Molecular Biology and Translational Science
|September 7, 2023
PubMed
Summary

Colony-forming units in liver (CFU-L) can reliably measure hepatocyte stemness and engraftment. This new assay offers a quantitative method for assessing liver stem cell function, unlike current methods for hepatocyte-like cells.

Keywords:
Hepatocyte-like cellschimeric micecolony-forming units in spleendrug-induced liver injuryextramedullary erythropoiesishematopoietic stem cellshepatic stem cellhuman pluripotent stem cells

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Area of Science:

  • Hepatology
  • Stem Cell Biology
  • Regenerative Medicine

Background:

  • Hepatocyte-like cells (HLCs) from human pluripotent stem cells (PSCs) show promise but have poor in vivo engraftment and function.
  • Current differentiation protocols have not overcome the engraftment limitations of HLCs in liver injury models.
  • Organ-derived hepatocytes demonstrate reproducible colony formation in vivo, unlike HLCs.

Purpose of the Study:

  • To propose and validate a novel quantitative assay for assessing hepatocyte stemness, engraftment, and functionality.
  • To introduce the concept of "colony-forming units in liver" (CFU-L) as a reliable indicator.
  • To establish CFU-L as a measure analogous to hematopoietic stem cell "colony-forming units in spleen" (CFU-s).

Main Methods:

  • Comparison of in vivo engraftment and colony formation between HLCs and organ-derived hepatocytes in a mouse liver injury model.
  • Hypothesizing CFU-L as a functional stemness assay based on observed colony formation.
  • Leveraging single-cell origin evidence (uniform gene inactivation) to support CFU-L as a quantitative measure.

Main Results:

  • Hepatocyte-like cells failed to engraft in the mouse liver injury model.
  • Organ-derived hepatocytes consistently formed colonies in the same model.
  • The concept of CFU-L is proposed as a direct measure of in vivo engrafting and proliferating hepatocytes.

Conclusions:

  • Colony-forming units in liver (CFU-L) represent a robust assay for quantifying hepatocyte stemness and in vivo engraftment potential.
  • CFU-L offers a standardized method for evaluating the functional capacity of hepatocytes, crucial for cell-based therapies.
  • This approach provides a new paradigm for stem cell assessment in liver research, mirroring established methods in hematopoiesis.