Stromal cell-independent differentiation of human cord blood CD34+CD38− lymphohematopoietic progenitors toward B cell lineage
| Autor: | Jun-ichi Ohkawara, Satoko Nakajo, Norihiro Sato, Miki Yamaguchi, Mitsuhiro Fujihara, Yoshihiro Yoshikawa, Sadayoshi Sekiguchi, M Kasai, Fumiya Hirayama, Makoto Kanai, Kenji Ikebuchi |
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| Rok vydání: | 2000 |
| Předmět: |
Cancer Research
Myeloid Cellular differentiation Gene Rearrangement B-Lymphocyte Heavy Chain Antigens CD34 Stem cell factor Biology Polymerase Chain Reaction Granulocyte Colony-Stimulating Factor medicine Humans Cell Lineage RNA Messenger Progenitor cell B cell B-Lymphocytes Stem Cell Factor Immunoglobulin mu-Chains Interleukin-6 Infant Newborn Cell Differentiation Hematology Fetal Blood Hematopoietic Stem Cells Molecular biology Recombinant Proteins Hematopoiesis Haematopoiesis medicine.anatomical_structure Oncology Cell culture Immunology Interleukin-3 Stromal Cells Stem cell Biomarkers |
| Zdroj: | Leukemia. 14:727-734 |
| ISSN: | 1476-5551 0887-6924 |
| Popis: | To study the cytokine regulation of early stages of human B-lymphopoiesis, we developed a stroma-free two-step culture system. Single human cord blood CD34+CD38- cells were individually cultured by micromanipulation with interleukin (IL)-3, stem cell factor (SCF), fIt3 ligand (FL), IL-6 and granulocyte colony-stimulating factor (G-CSF). About 10% of the cells formed primary colonies, which were individually tested for myeloid and B-lymphoid potentials by reculturing aliquots of the primary colony cells into secondary myeloid and B-lymphoid cultures. One third of the primary colonies proved capable of differentiation into CD19+IgM+ cells, as well as into myeloid lineage cells. RT-PCR analyses revealed that some cells in the primary culture had already matured to express B cell-specific transcripts. Thus, the combination of IL-3, SCF, FL, IL-6 and G-CSF supported the differentiation of CD34+CD38- lymphohematopoietic progenitors toward B cell lineage in addition to myeloid lineages. Screening of cytokines to identify the minimum requirement of cytokines in the primary culture revealed that IL-3 and SCF were essential and that the addition of FL, and to a lesser extent IL-6 or G-CSF, to the combination of IL3 and SCF remarkably enhanced the primary colony formation and the generation of CD19+ cells in the secondary B-lymphoid culture. |
| Databáze: | OpenAIRE |
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