Full reconstitution of hematopoietic system by murine umbilical cord blood.
BACKGROUND
Murine umbilical cord blood cells (UCBCs) were studied for their ability to reconstitute the hematopoietic system.


METHODS
On average, 150 microL of cord blood per fetus containing 1.2 to 2 x 10(4) nucleated cells were collected from day 18.5 fetal umbilical cord, and 3 to 6 x 10(3) cells per fetus were obtained after separation by gradient centrifugation.


RESULTS
Although lineage marker-, c-Kit+, and Sca-1+ cells were detectable among UCBCs, cells designated to be in the side population (SP) by Hoechst 33342 staining were hardly detectable within this population; the frequency of cells of this phenotype was less than 1 of 105. Instead, the lineage marker-, c-Kit+, and Sca-1- population contained a considerable number of SP cells. Nevertheless, UCBCs obtained from fetuses of green fluorescent protein-transgenic mice successfully reconstituted the blood cells of lethally irradiated recipients. Fluorescent cells could be readily detected in every blood cell lineage and among immature cell populations. Furthermore, fluorescent SP cells sorted from the recipient bone marrow cells could also reconstitute the blood cells in the secondary recipients, indicating that UCBCs also replenished bone marrow stem cells.


CONCLUSIONS
Murine UCBC could fully reconstitute the hematopoietic system of lethally irradiated recipients including hematopoietic stem cells in bone marrow.
Migishima
Fujio
F
Department of Obstetrics and Gynecology, Kitasato University School of Medicine, Sagamihara, Japan.
Oikawa
Atsuhiko
A
Kondo
Shunzo
S
Ema
Hideo
H
Morita
Yohei
Y
Nakauchi
Hiromitsu
H
Yokoyama
Minesuke
M
Song
Si-Young
SY
Nishijima
Masahiro
M
Okabe
Masaru
M
Shinohara
Nobukata
N
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis; D001562:Benzimidazoles; D019070:Cell Lineage; D036101:Cord Blood Stem Cell Transplantation; D005260:Female; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology*; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis; D018183:Transplantation Chimera; D014916:Whole-Body Irradiation
D000950:Antigens, Ly; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D008164:Luminescent Proteins; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D049452:Green Fluorescent Proteins; D019009:Proto-Oncogene Proteins c-kit; C017807:bisbenzimide ethoxide trihydrochloride