PCR amplification of short tandem repeat sequences allows serial studies of chimaerism/engraftment following BMT in rodents.
Animal models of bone marrow transplantation (BMT) allow evaluation of new experimental treatment strategies. One potential strategy involves the treatment of donor marrow with ultra-violet B light to allow transplantation across histocompatibility boundaries without an increase in graft rejection or graft-versus-host disease. A major requirement for a new experimental protocol, particularly if it involves manipulation of the donor marrow, is that the manipulated marrow gives rise to long-term multilineage engraftment. DNA based methodologies are now routinely used by many centres to evaluate engraftment and degree of chimaerism post-BMT in humans. We report the adaptation of this methodology to the serial study of engraftment in rodents. Conditions have been defined which allow analysis of serial tail vein samples using PCR of short tandem repeat sequences (STR-PCR). These markers have been used to evaluate the contribution of ultraviolet B treated marrow to engraftment following BMT in rodents without compromising the health of the animals under study. Chimaerism data from sequential tail vein samples and bone marrow from selected sacrificed animals showed excellent correlation, thus confirming the validity of this approach in analysing haemopoietic tissue. Thus the use of this assay may facilitate experimental studies in animal BMT.
O'Neill
P A
PA
Department of Haematology and Oncology, Sir Patrick Duns Research Labs, St James's Hospital, Dublin, Ireland.
Lawler
M
M
Pullens
R
R
Kloosterman
T
T
Hudson
J
J
Martens
A C
AC
Hendrikx
P J
PJ
Gowing
H
H
Byrne
C
C
Hagenbeek
A
A
Pamphilon
D H
DH
McCann
S R
SR
D000818:Animals; D001483:Base Sequence; D016026:Bone Marrow Transplantation / Q000473:pathology*; D006085:Graft Survival / Q000235:genetics; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000528:radiation effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011828:Radiation Chimera / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D014466:Ultraviolet Rays