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In Vitro Differentiation of Murine Sca-1⁺Lin^– Cells into Myeloid, B Cell and T Cell Lineages

Department of Transfusion Medicine, Hyogo College of Medicine, Nishinomiya City, Japan

Key Words. T cell differentiation • Hematopoietic progenitors • B cell differentiation

Dr. Hiroshi Hara, Department of Transfusion Medicine, Hyogo College of Medicine, 1-1, Mukogawa-cho, Nishinomiya City, Hyogo, 663, Japan.

Hematopoietic progenitor cells were shown to be capable of differentiating into myeloid, B cell and T cell lineages. We used a two-step culture system in which enriched murine hematopoietic progenitors in bone marrow were first plated in viscid culture medium containing methylcellulose, erythropoietin (EPO), stem cell factor (SCF) and interleukin (IL)-7. One thousand enriched murine marrow cells formed 53.5 ± 12.1 (mean ± SD) primary colonies. Cells from a single blast colony were separated into two aliquots and replated in secondary methylcellulose cultures containing SCF and IL-7 for B cell lineage and SCF, IL-3, G-CSF, GM-CSF and EPO for myeloid lineage. Next, cells from five to ten primary blast colonies were cultured again in embryonal thymus (25 Gy irradiated). One aliquot of blast colonies in a primary culture contained four colony forming units (CFU) of granulocytes, erythroblasts, macrophages and megakaryocytes, eight CFU-granulocytes and macrophages, and 28 BFU-E in a representative secondary myeloid culture. Another aliquot formed a few B cell colonies (2-10) in a secondary B cell culture. B lymphoid colonies were composed of blast-like cells with B-220 antigen. T cells in a secondary T cell culture consisted of 16% L3T4⁺, 16% CD8⁺ and 11% CD3⁺ of bone marrow origin in the thymus. From these results, we concluded that cells in the primary colonies from Sca-1⁺Lin^– hematopoietic stem cells could differentiate into B cell, T cell and myeloid lineages.

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