Comparison of strategies to detect and quantitate uniquely marked cells in intra- and inter-species hemopoietic chimeras

M. G. Pallavicini, R. G. Langlois, M. Reitsma, Mark L Gonzalgo, D. Sudar, T. Montoya, H. U. Weier, S. Haendel

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Evaluation of the outcome of successful bone marro transplantation and in-depth studies of transplantation biology rely increasingly upon detection and enumeration of donor hemopoietic cells in the transplanted recipients. The ability to detect and enumerate low levels of donor engraftment in interphase cell subpopulations in hemopoietic chimeras is particularly important for studies of mixed lineage chimerism, early relapse manifestations, and engraftment of subpopulations present at low frequency. We describe and compare the sensitivity and specificity of DNA-based detection strategies (fluorescence in situ hybridization, in vitro DNA amplification using the polymerase chain reaction) and flow cytometric analysis of cell surface markers to detect cells carrying marker DNA or proteins in syngeneic (mouse-to-mouse) and xenogeneic (mouse-to-human, monkey, sheep) backgrounds. DNA-based detection strategies offer advantages of rapid analysis and enumeration of target cell frequencies with detection sensitivities approximating 10-4. The sensitivity of immunofluorescence-linked flow cytometric-based detection of nucleated leukocytes approached 10-3, whereas flow cytometric-based detection of fixed human erythrocytes was feasible at cell frequencies of 10-5. Data described in this manuscript should facilitate selection of appropriate methodologies for assessment of hemopoietic chimerism following transplantation.

Original languageEnglish (US)
Pages (from-to)356-367
Number of pages12
JournalCytometry
Volume13
Issue number4
DOIs
StatePublished - 1992
Externally publishedYes

Fingerprint

Chimerism
DNA
Transplantation
Bone Transplantation
Interphase
Fluorescence In Situ Hybridization
Genetic Markers
Haplorhini
Fluorescent Antibody Technique
Sheep
Leukocytes
Erythrocytes
Recurrence
Sensitivity and Specificity
Polymerase Chain Reaction
Proteins
In Vitro Techniques

Keywords

  • Immunofluorescence
  • Markers
  • Polymerase chain reaction
  • Transplantation

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Comparison of strategies to detect and quantitate uniquely marked cells in intra- and inter-species hemopoietic chimeras. / Pallavicini, M. G.; Langlois, R. G.; Reitsma, M.; Gonzalgo, Mark L; Sudar, D.; Montoya, T.; Weier, H. U.; Haendel, S.

In: Cytometry, Vol. 13, No. 4, 1992, p. 356-367.

Research output: Contribution to journalArticle

Pallavicini, MG, Langlois, RG, Reitsma, M, Gonzalgo, ML, Sudar, D, Montoya, T, Weier, HU & Haendel, S 1992, 'Comparison of strategies to detect and quantitate uniquely marked cells in intra- and inter-species hemopoietic chimeras', Cytometry, vol. 13, no. 4, pp. 356-367. https://doi.org/10.1002/cyto.990130405
Pallavicini, M. G. ; Langlois, R. G. ; Reitsma, M. ; Gonzalgo, Mark L ; Sudar, D. ; Montoya, T. ; Weier, H. U. ; Haendel, S. / Comparison of strategies to detect and quantitate uniquely marked cells in intra- and inter-species hemopoietic chimeras. In: Cytometry. 1992 ; Vol. 13, No. 4. pp. 356-367.
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