Residues 39-56 of Stem Cell Factor Protein Sequence Are Capable of Stimulating the Expansion of Cord Blood CD34+ Cells.
BACKGROUND
Stem cell factor (SCF) can stimulate hematopoietic stem cell (HSC) expansion; however, the specific structural region(s) of SCF protein that are critical for this function are still unknown. A novel monoclonal antibody (named 23C8) against recombinant human SCF (rhSCF) was previously found to inhibit the ability of rhSCF to enhance HSC expansion, making it possible to identify the relevant active region to HSC.


METHODS
Eleven polypeptides were synthesized, which were designed to cover the full-length of rhSCF, with overlaps that are at least 3 amino acids long. ELISA was used to identify the polypeptide(s) that specifically react with the anti-SCF. The effects of the synthetic polypeptides on human HSC expansion, or on the ability of the full-length rhSCF to stimulate cell proliferation, were evaluated ex vivo. Total cell number was monitored using hemocytometer whereas CD34+ cell number was calculated based on the proportion determined via flow cytometry on day 6 of culture.


RESULTS
Of all polypeptides analyzed, only one, named P0, corresponding to the SCF protein sequence at residues 39-56, was recognized by 23C8 mAb during ELISA. P0 stimulated the expansion of CD34+ cells derived from human umbilical cord blood (UCB). Addition of P0 increased the numbers of total mononucleated cells and CD34+ cells, by ~2 fold on day 6. P0 also showed partial competition against full-length rhSCF in the ex vivo cell expansion assay.


CONCLUSIONS
Residues 39-56 of rhSCF comprise a critical functional region for its ability to enhance expansion of human UCB CD34+ cells. The peptide P0 is a potential candidate for further development as a synthetic substitute for rhSCF in laboratory and clinical applications.
Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China.
Bin
B
Shen
Biopharmagen Corp, Suzhou, China.
Wenhong
W
Jiang
Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China.
Jie
J
Fan
Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China; Department of Environmental Medicine, New York University Langone Medical Center, Tuxedo, New York, United States of America.
Wei
W
Dai
Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China; College of Nanoscale Science, SUNY Polytechnic Institute, Albany, New York, United States of America.
Xinxin
X
Ding
Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China.
Yongping
Y
Jiang
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D018952:Antigens, CD34 / Q000276:immunology; D002453:Cell Cycle / Q000235:genetics; D049109:Cell Proliferation; D005312:Fetal Blood / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D019089:Stem Cell Factor / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D019089:Stem Cell Factor