title: In vitro production of functional immune cells derived from human hematopoietic stem cells.
abstract: Hematopoietic stem cells (HSC) from cord blood are potentially high sources for transplantation due to their low immunogenicity and the presence of the multipotent cells. These cells are capable of differentiating to produce various lineages of blood cells under specific conditions. We have enriched highly purified CD34(+) cells from cord blood, determined in vitro growth of the cells in culture systems in the absence (condition A) or presence of GM-CSF and G-CSF (condition B), and determined the profile of immune cells during the period of cultivation by using flow cytometry. PhytohemagglutininA (PHA) was used as a mitogen to stimulate T lymphocytes derived from hematopoietic stem cells. GM-CSF and G-CSF prolonged the survival of the growing cells and also maintained expansion of cells in blastic stage. By day 12 of cultivation, when cell numbers peaked, various types of immune cells had appeared (CD14(+) cells, CD40(+)HLA-DR(+) cells, CD3(+)CD56(+) cells, CD19(+) cells, CD3(+)CD4(+) cells, CD3(+)CD8(+)cells and CD3-CD56(+)). A significantly higher percentage of monocytes (p = 0.002) were observed under culture with GM-CSF, G-CSF when compared with culture without GM-CSF, G-CSF. In addition, T lymphocytes derived from HSC responded to 50 µg/ml of PHA. This is the first report showing the complete differentiation and proliferation of immune cells derived from CD34(+) HSC under in vitro culture conditions. Lymphocytes, monocytes, dendritic cells and polymorph nuclear cells derived from HSC in vitro are unique, and thus may benefit various studies such as innate immunity and pathophysiology of immune disorders.
authors.affiliation: Department of Pathobiology, Faculty of Science, Mahidol University, Thailand.
authors.forename: Witchuda
authors.initials: W
authors.lastname: Payuhakrit
authors.affiliation: Faculty of Science and Technology, Suan Dusit Rajabhat University, Thailand.
authors.forename: Tasanee
authors.initials: T
authors.lastname: Panichakul
authors.affiliation: Department of Pathobiology, Faculty of Science, Mahidol University, Thailand.
authors.forename: Natthawut
authors.initials: N
authors.lastname: Charoenphon
authors.affiliation: Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Thailand.
authors.forename: Panus
authors.initials: P
authors.lastname: Chalermsaenyakorn
authors.affiliation: Department of Obstetrics and Gynaecology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Thailand.
authors.forename: Adithep
authors.initials: A
authors.lastname: Jaovisidha
authors.affiliation: Research Center, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Thailand.
authors.forename: Chokdee
authors.initials: C
authors.lastname: Wongborisuth
authors.affiliation: Department of Pathobiology, Faculty of Science, Mahidol University, Thailand; Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Thailand; Center for Emerging and Neglected Infectious Diseases, Mahidol University, Thailand.
authors.forename: Rachanee
authors.initials: R
authors.lastname: Udomsangpetch
keywords: hematopoietic stem cells; lymphocyte maturation; myelopoietic cells; stem cells