Ex vivo expanded murine bone marrow cells with a multiple cytokine cocktail retain long-term hematopoietic reconstitution potentials.
BACKGROUND
Although ex vivo expansion of bone marrow (BM) cells has been proposed as an effective method for the early recovery from pancytopenia in patients with bone marrow stem cell transplantation (BMT), the expansion potential and the long-term reconstitution capability of such BM cells is still controversial. We describe here a multiple cytokine medium (MCM) containing major hematopoietic stimulation factors and conditioned medium from PHA-stimulated murine spleen cells that permits the expansion of BM cells with long-term hematopoietic reconstitution capacity.


METHODS
Male murine BM cells were expanded in MCM for 4 to 14 days and injected into lethally irradiated syngeneic female mice. The mice were maintained for 18 months after transplantation for evaluation of hematopoietic reconstitution.


RESULTS
The expanded cells contained pluripotent hematopoietic stem cells and lineage committed progenitors as well as terminally differentiated cells. They permitted full recovery of lethally irradiated mice in both early and late stages in same numbers equivalent to that of unexpanded cells. More than 80% of the progenitor cells were donor originated after 18 months. Expanded cells were able to be transduced with a retroviral vector expressing Beta-galactosidase, and continued to express the marker following BMT.


CONCLUSIONS
With the use of MCM, the quantity of donor cells from BM and other sources might be greatly reduced. Ex vivo expanded BM cells might also facilitate gene manipulation in vitro by retroviral vectors.
Yang
Quan-En
QE
Laboratory of Cell and Molecular Structure, SAIC/Frederick, Inc., National Cancer Institute at Frederick, Frederick, MD 21702, USA. yangq@dtpax2.ncifcrf.gov
Fong
Steven E
SE
Li
Kungui
K
Gonda
Matthew A
MA
Tobin
Gregory J
GJ
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D016026:Bone Marrow Transplantation / Q000502:physiology; D018929:Cell Culture Techniques; D002455:Cell Division / Q000187:drug effects; D003114:Colony-Forming Units Assay; D016207:Cytokines / Q000494:pharmacology*; D017931:DNA Primers; D006410:Hematopoiesis / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010835:Phytohemagglutinins / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D014998:Y Chromosome
D016207:Cytokines; D017931:DNA Primers; D010835:Phytohemagglutinins