c-kit<low Pluripotent hemopoietic stem cells form CFU-S on day 16.
Using Ly5 congenic mice, we characterized the early differentiation step of pluripotent hemopoietic stem cells. Lineage- (Lin-)/CD71- cells in the bone marrow cells were separated into major histocompatibility complex (MHC) class I(high)/c-kit(low) and MHC class I(high)/c-kit<low populations from C57BL/6 Ly5.1 male mice. These two populations (1,000 cells) were transplanted into lethally irradiated (5.5 Gy x 2) C57BL/6 Ly5.2 female mice. Colony-forming unit in spleen (CFU-S) assays were carried out on days 10, 12, 14, 16, and 20. In the mice that received c-kit(low) cells, CFU-S were first detected on day 12, and the CFU-S counts gradually increased. In contrast, no visible colony was detected until day 14 in the mice that received c-kit<low cells; CFU-S were first observed on day 16. Donor-derived (Ly5.1+) cells, such as B cells, T cells, and myeloid cells, were detected by fluorescence-activated cell sorter analyses, and donor-derived erythroid cells were detected by polymerase chain reaction analyses using Y-chromosome-specific primers. Donor-derived cells in the recipients of c-kit(low) cells were detected in the spleen, bone marrow, and peripheral blood on day 12 after transplantation, while they were detected on day 16 in the mice that received c-kit<low cells. Therefore, c-kit<low cells have the capacity not only to form CFU-S on day 16 but also to reconstitute the recipients with donor-derived hematolymphoid cells 16 days after transplantation.
Lian
Z
Z
First Department of Pathology, Kansai Medical University, Moriguchi City, Osaka, Japan.
Feng
B
B
Sugiura
K
K
Inaba
M
M
Yu
C
C
Jin
T
T
Fan
T
T
Cui
Y
Y
Yasumizu
R
R
Toki
J
J
Adachi
Y
Y
Hisha
H
H
Ikehara
S
S
D000818:Animals; D015704:CD4 Antigens / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D003114:Colony-Forming Units Assay; D015672:Erythroid Precursor Cells / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D005472:Fluorouracil / Q000494:pharmacology; D006098:Granulocytes / Q000276:immunology; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D017493:Leukocyte Common Antigens / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D008297:Male; D051379:Mice; D020297:Mice, Congenic; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000737:chemistry / Q000166:cytology*; D019089:Stem Cell Factor / Q000097:blood / Q000494:pharmacology*; D013997:Time Factors
D015704:CD4 Antigens; D016827:CD8 Antigens; D019089:Stem Cell Factor; D017493:Leukocyte Common Antigens; D005472:Fluorouracil