Avoiding pitfalls in bone marrow engraftment analysis: a case study highlighting the weakness of using buccal cells for determining a patient's constitutional genotype after hematopoietic stem cell transplantation |
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| Authors: | Hanna Rennert Debra GB Leonard Melissa Cushing Carmen Azurin Tsiporah Shore |
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| Institution: | 1. Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, New York, USA;2. Department of Medicine, Weill Cornell Medical College, New York, New York, USA;1. Chemical & Biomedical Engineering, FAMU-FSU College of Engineering, The Florida State University, Tallahassee, Florida, USA;2. The National High Magnetic Field Laboratory, The Florida State University, Tallahassee, Florida, USA;1. National Marrow Donor Program, Minneapolis, MN, USA;2. Manipal Hospital, Bangalore, India;3. Rotary Bangalore-TTK Blood Bank, Bangalore Medical Services Trust, Bangalore, India;4. Narayana Hrudayalaya, Bangalore, India;5. Jeevan Stem Cell Bank, Chennai, India;6. Tata Memorial Hospital, Mumbai, India;7. Post Graduate Institute of Medical Education and Research, Chandigarh, India;8. Gujarat Cancer and Research Institute, Civil Hospital Campus, Ahmedabad, India;9. DATRI, Chennai, India;10. Histogenetics, New York, NY, USA;11. Marrow Donor Registry (India), Mumbai, India;12. Christian Medical College, Vellore, India;13. Division of Hematology, Oncology and Transplantation, University of Minnesota, MN, USA;2. Laboratory of Cell and Genetic Therapy, University of Liège, Liège, Belgium;3. Department of Pathology, University of Liège, Liège, Belgium;4. Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium;1. Berlin-Brandenburg Center for Regenerative Therapies, Charité University Medicine, Campus Virchow, Berlin, Germany;2. Institute of Medical Immunology, Charité University Medicine, Campus Virchow, Berlin, Germany;3. Department of Nephrology and Internal Intensive Care, Charité University Medicine, Campus Virchow, Berlin, Germany;4. Institute for Regenerative Cures, University of California Davis, Sacramento, California, USA |
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| Abstract: | Background aimsAn accurate and reliable assessment of bone marrow engraftment (BME) after hematopoietic stem cell transplantation (HSCT) is based on the ability to distinguish between recipient and donor cells at selected polymorphic short tandem repeat (STR) DNA loci. Buccal cells are an important source of DNA for determining the recipient's constitutional genotype, particularly in patients transplanted before the STR evaluation.MethodsGenomic DNA was extracted from the recipient buccal cells and from isolated CD3+ (T-cell lymphocyte) and CD33+ (myelocyte) cells after HSCT. BME analysis was performed using a STR-based polymerase chain reaction amplification method followed by fragment-size analysis for assessing the recipient-derived or donor-derived composition of cell lineage-specific peripheral blood DNA.ResultsWe identified three cases of complete buccal epithelial cell engraftment after HSCT detected by BME analysis, potentially leading to misinterpretation of testing results if these cells were used as the sole source for determining the recipient's genotype.ConclusionsThese cases suggest that complete engraftment of buccal epithelial cells may be a common finding in patients receiving HSCT, drawing attention to important issues such as the type of samples used for determining a patient's constitutional genotype that may confound testing results. This study also highlights the need for careful interpretation of the BME testing results in the context of the clinical findings. |
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