Collection and analysis of hematopoietic progenitor cells from cynomolgus macaques (Macaca fascicularis): assessment of cross-reacting monoclonal antibodies.
Previous studies have shown that hematopoietic progenitor cells can be isolated from human or nonhuman primate bone marrow (BM) cells. In the present study, we studied the cross-reactivity of 13 anti-human CD34, two anti-human c-Kit, and one anti-human CD133 monoclonal antibodies (mAbs) with cynomolgus macaque (Macaca fascicularis) BM cells, using flow cytometric analysis, cell enrichment, and clonogenic assay. Among the 13 anti-human CD34 mAbs assessed, six cross-reacted as previously reported by other groups. However, only three of these six mAbs (clones 561, 563, and 12.8) recognized cynomolgus CD34+ cells that formed progenitor colonies when grown in methylcellulose culture. Similarly, of the two anti-human c-Kit mAbs (clones NU-c-kit and 95C3) that were previously reported to cross-react with cynomolgus BM cells, only one (clone NU-c-kit) resulted in a similar outcome. The anti-human CD133 mAb (clone AC133) also cross-reacted with cynomolgus BM cells, although these cells did not give rise to colonies when grown in culture. These results suggest that antibodies that cross-react with nonhuman primate cells may not identify the hematopoietic cells of interest. In addition, while the CD34 mAb (clone 561) results in the selection of hematopoietic progenitor cells of all lineages when assessed in methylcellulose culture, the c-Kit(high) fraction (NU-c-kit) exclusively identifies erythroid-specific progenitor cells after growth in culture. It is important to consider these findings when selecting cross-reacting mAbs to identify cells of hematopoietic lineages in macaque species.
Shibata
Hiroaki
H
Tsukuba Primate Center, National Institute of Infectious Diseases, Ibaraki, Japan.
Hanazono
Yutaka
Y
Ageyama
Naohide
N
Nagashima
Takeyuki
T
Ueda
Yasuji
Y
Hasegawa
Mamoru
M
Ozawa
Keiya
K
Yoshikawa
Yasuhiro
Y
Terao
Keiji
K
D000071916:AC133 Antigen; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology*; D002469:Cell Separation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D003429:Cross Reactions; D005260:Female; D005434:Flow Cytometry; D006023:Glycoproteins / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology*; D006801:Humans; D008252:Macaca fascicularis / Q000276:immunology*; D008297:Male; D010455:Peptides / Q000276:immunology*; D019009:Proto-Oncogene Proteins c-kit / Q000276:immunology*; D013045:Species Specificity
D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018952:Antigens, CD34; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D019009:Proto-Oncogene Proteins c-kit