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Donor-derived hepatocytes in human hematopoietic cell transplant recipients: evidence of fusion

  • David MyersonEmail author
  • Rachael K. Parkin
Original Article

Abstract

Reconstitution of hepatocytes by hematopoietic stem cells—a phenomenon which occurs in rodents under highly selective conditions—results from infrequent fusion between incoming myelomonocytes and host hepatocytes, with subsequent proliferation. Human hematopoietic stem cell transplant recipients have been little studied, with some support for transdifferentiation (direct differentiation). We studied routinely obtained autopsy liver tissue of four female hematopoietic cell transplant recipients with male donors, using a highly specific conjoint immunohistochemistry in situ hybridization light microscopic technique. Hepatocyte nuclei were identified by cytokeratin (Cam5.2) staining and evaluated for X and Y chromosome content. Over 1.6 million hepatocytes were assessed for rare instances of donor origin, revealing a Y chromosome in 67. Mixed tetraploids (XXXY) and their nuclear truncation products (XXY, XY, Y) were directly demonstrated, with no detection of the male tetraploids (XXYY) that may result from transdifferentiation with subsequent tetraploidization, nor their unique truncation products (XYY, YY), implicating fusion as the mechanism. To determine whether it is the sole mechanism, we modeled the chromosome distribution based on the same probability of detection of each X chromosome, deriving parameters of sensitivity and female tetraploidy by best fit. We then hypothesized that the distribution of Y chromosome–containing cells could be predicted by a similar model. After modification to account for “clumpy” Y chromosomes, the observed results were in accord with the predicted results (p = 0.6). These results suggest that all the Y-containing cells, including apparent XY cells, derive from mixed tetraploids, consistent with fusion as the sole mechanism.

Keywords

Liver regeneration Stem cells Progenitor cells In situ hybridization Hematopoietic stem cell transplant 

Notes

Acknowledgments

We thank Dr. Ted A. Gooley for statistics advice.

Contributions

DM conceived and designed the study; analyzed the data; and wrote, edited, and reviewed the manuscript. RKP acquired the data and edited and reviewed the manuscript. Both authors read and approved the final manuscript.

Funding

This work was supported in part by grants from the National Institutes of Health CA18029, CA15074, and HL36444.

Compliance with ethical standards

The study was performed under a protocol approved by the Institutional Review Board of the Fred Hutchinson Cancer Research Center.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Clinical Research DivisionFred Hutchinson Cancer Research CenterSeattleUSA
  2. 2.Department of PathologyUniversity of WashingtonSeattleUSA

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