Hematopoietic stem cell origin of human fibroblasts: Cell culture studies of female recipients of gender-mismatched stem cell transplantation and patients with chronic myelogenous leukemia

Shirai, Keisuke; Sera, Yasuhiko; Bulkeley, William; Mehrotra, Meenal; Moussa, Omar; LaRue, Amanda C.; Watson, Dennis K.; Stuart, Robert K.; Lazarchick, John; Ogawa, Makio

doi:10.1016/j.exphem.2009.09.008

« PreviousExperimental Hematology
Volume 37, Issue 12 , Pages 1464-1471, December 2009

Hematopoietic stem cell origin of human fibroblasts: Cell culture studies of female recipients of gender-mismatched stem cell transplantation and patients with chronic myelogenous leukemia

Keisuke Shirai
- Department of Medicine, Medical University of South Carolina, Charleston, SC., USA
Yasuhiko Sera
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
- Department of Veterans Affairs Medical Center, Charleston, SC., USA
William Bulkeley
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
Meenal Mehrotra
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
- Department of Veterans Affairs Medical Center, Charleston, SC., USA
Omar Moussa
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
Amanda C. LaRue
- Department of Veterans Affairs Medical Center, Charleston, SC., USA
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
Dennis K. Watson
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
Robert K. Stuart
- Department of Medicine, Medical University of South Carolina, Charleston, SC., USA
John Lazarchick
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
Makio Ogawa
- Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC., USA
- Department of Veterans Affairs Medical Center, Charleston, SC., USA
- Offprint request to: Makio Ogawa, M.D., Ph.D., Department of Veterans Affairs Medical Center, 109 Bee Street, Charleston, SC 29401

Received 23 September 2009; received in revised form 23 September 2009; accepted 23 September 2009. published online 28 September 2009.

Objective

Our series of studies using transplantation of single hematopoietic stem cells (HSCs) demonstrated that mouse fibroblasts/myofibroblasts are derived from HSCs. In order to determine the origin of human fibroblasts, we established a method for culturing fibroblasts from human peripheral blood (PB) mononuclear cells and studied fibroblasts from gender-mismatched HSC transplant recipients and patients with untreated Philadelphia chromosome–positive chronic myelogenous leukemia (CML).

Materials and Methods

We cultured PB cells from three female subjects who showed near-complete hematopoietic reconstitution from transplantation of granulocyte-colony stimulating factor–mobilized male PB cells and examined the resulting fibroblasts using fluorescent in situ hybridization for Y chromosome. Because the mobilized PB cells may contain mesenchymal stem cells, we could not determine the HSC or mesenchymal stem cell origin of the fibroblasts seen in culture. To further document the HSC origin of human fibroblasts, we next examined fibroblasts from two patients with untreated CML, a known clonal disorder of HSCs.

Results

All cultured fibroblasts from female recipients of male cells showed the presence of Y chromosome, indicating the donor origin of fibroblasts. Cultured fibroblasts from the CML patients revealed the presence of BCR-ABL translocation. This demonstration provided strong evidence for the HSC origin of human fibroblasts because CML is a clonal disorder of the HSC.

Conclusions

These studies strongly suggest that human fibroblasts are derived from HSCs. In addition, the results suggest that fibrosis seen in patients with CML may be a part of the clonal process.