The proposed studies will examine the effects of two novel hematopoietic growth factors - stem cell factor (SCF) and Rauscher factor - on the self- renewal and differentiation of highly enriched populations of murine and human hematopoietic stem cells (HSC). Recombinant purified rat SCF will be studied for its effect alone and with other growth factors such as IL-1, IL-3, IL-6, GM-CSF and erythropoietin (Epo) on the self renewal and differentiation of HSC isolated by physical means. Suspensions of HSC will be used in """"""""read out"""""""" assays for the generation of colony-forming cells (CFC). In vivo, murine HSC function will be assayed by day 12 spleen colony-formation and long-term marrow reconstitution. The effects of recombinant human HSC will be studied using highly enriched populations of human progenitor cells (from marrow and umbilical cord blood) separated by flow cytometry or derived from blast cell colonies. SCF will be tested alone and in combination with other hematopoietic growth factors for generation of CFU-Mix and unilineage CFC in read-out assays. Rauscher factor, a potentially novel growth factor which is not species- specific, is most easily detectable when hematopoietic CFC come into contact with irradiated Rauscher cells. Thus, Rauscher cells may form a matrix containing Rauscher factor which can be studied by adding enriched murine or human SCF in low numbers to an established feeder. All cultures will be conducted under serum-deprived conditions to eliminate factors or co-factors provided in fetal bovine serum. These studies should provide insight into the mechanisms of action of these factors and the range of progenitor cells acted upon. The results also may allow a basis for the use of combinations of these and other factors to amplify HSC numbers appropriate for transplantation or quantities of differentiated cells appropriate for transfusion.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL046524-02
Application #
3365641
Study Section
Special Emphasis Panel (SRC (JD))
Project Start
1991-05-01
Project End
1995-02-28
Budget Start
1992-03-01
Budget End
1993-02-28
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
New York Blood Center
Department
Type
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10065
Migliaccio, G; Baiocchi, M; Adamson, J W et al. (1996) Isolation and biological characterization of two classes of blast-cell colony-forming cells from normal murine marrow. Blood 87:4091-9
Eddleman, K A; Chervenak, F A; George-Siegel, P et al. (1996) Circulating hematopoietic stem cell populations in human fetuses: implications for fetal gene therapy and alterations with in utero red cell transfusion. Fetal Diagn Ther 11:231-40
Migliaccio, G; Baiocchi, M; Hamel, N et al. (1996) Circulating progenitor cells in human ontogenesis: response to growth factors and replating potential. J Hematother 5:161-70
Ashany, D; Elkon, K B; Migliaccio, G et al. (1995) Functional characterization of lymphoid cells generated in serum-deprived culture stimulated with stem cell factor and interleukin 7 from normal and autoimmune mice. J Cell Physiol 164:562-70
Durand, B; Migliaccio, G; Yee, N S et al. (1994) Long-term generation of human mast cells in serum-free cultures of CD34+ cord blood cells stimulated with stem cell factor and interleukin-3. Blood 84:3667-74
Migliaccio, A R; Migliaccio, G; Durand, B et al. (1993) The generation of colony-forming cells (CFC) and the expansion of hematopoiesis in cultures of human cord blood cells is dependent on the presence of stem cell factor (SCF). Cytotechnology 11:107-13
Migliaccio, A R; Migliaccio, G; Mancini, G et al. (1993) Induction of the murine ""W phenotype"" in long-term cultures of human cord blood cells by c-kit antisense oligomers. J Cell Physiol 157:158-63
Nicolis, S; Ottolenghi, S; Papayannopoulou, T et al. (1993) Dependence for the proliferative response to erythropoietin on an established erythroid differentiation program in a human hematopoietic cell line, UT-7. Exp Hematol 21:665-70
Durand, B; Eddleman, K; Migliaccio, A R et al. (1993) Long-term generation of colony-forming cells (CFC) from CD34+ human umbilical cord blood cells. Leuk Lymphoma 11:263-73
Migliaccio, G; Migliaccio, A R; Druzin, M L et al. (1992) Long-term generation of colony-forming cells in liquid culture of CD34+ cord blood cells in the presence of recombinant human stem cell factor. Blood 79:2620-7

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