A combination of stem cell factor and granulocyte colony-stimulating factor enhances the growth of human progenitor B cells supported by murine stromal cell line MS-5

Abstract

We have developed a long-term culture system using the murine bone marrow stromal cells MS-5 to support the growth of progenitor B cells with CD34-, CD10 + , CD19 + , and cyto-plasmic ? chain (C ?)-negative surface phenotype from human CD34 + cells purified from umbilical cord blood (CB). When 10 3 CB CD34 + cells/well were seeded on MS-5 stromal cells at the beginning of culture in the absence of exogenously added cytokines, progenitor B cells first appeared after 14 days, and the maximal cell production was achieved during the 6th week of culture. Intriguingly, the addition of recombinant human stem cell factor (rhSCF) and granulocyte colony-stimulating factor (rhG-CSF), but not rhIL-7, strikingly enhanced the growth of progenitor B cells from CB CD34 + population cultured on MS-5 stromal cells. The culture of progenitor B cells could be maintained until the 6th week of culture when some cells were revealed to have a C ? + phenotype, and a small number of cells had immunoglobu-lin ? chain on their cell surface in the presence of both rhSCF and rhG-CSF. When CD34 + cells were cultured physically separated from the stromal layer by membrane, supportive effects of MS-5 stromal cells for the growth of progenitor B cells were not observed. These results suggest that the present culture system could generate progenitor B cells to proliferate from CB CD34 + cells, that some of these progenitor B cells could differentiate into immature B cells in conjunction with rhSCF and rhG-CSF, and that a species-cross-reactive membrane-bound factor(s), which stimulates early human B lymphopoiesis, may exist in MS-5 stromal cells. Further studies are required to investigate the mechanism how rhG-CSF acts on progenitor B cells to allow their proliferation and differentiation.

Adiponectin, a Fat Cell Product, Influences the Earliest Lymphocyte Precursors in Bone Marrow Cultures by Activation of the Cyclooxygenase-Prostaglandin Pathway in Stromal Cells

“…Human CB CD34 ϩ CD38 Ϫ cells were cultured with MS-5 cells in ␣-MEM medium containing 10% FCS, 100 ng/ml recombinant human SCF, and 10 ng/ml recombinant human G-CSF (34). They were directly sorted into wells of 96-well flat-bottom plates preseeded with MS-5 cells, and then cultured for 6 wk.…”

“…Fortunately, the murine MS-5 stromal cell clone used in the experiments previously described also supports formation of human B lineage lymphocytes in culture (34). The hematopoietic stem cell-enriched CD34 ϩ CD38 Ϫ fraction was isolated from umbilical CB, and two cell concentrations were used to initiate cocultures on MS-5 monolayers (Table I).…”

Effect of a selective Abl tyrosine kinase inhibitor, STI571, on in vitro growth of BCR-ABL-positive acute lymphoblastic leukemia cells

“…Campana and colleagues developed an assay system by employing co-culture with human bone marrow-derived stromal cells, which provided reproducible growth of leukemic B-precursor cells from children with ALL. [22][23][24] Nishihara et al 25 recently reported a similar but xenogenic co-culture system for normal pro-B cells with the mouse bone marrow-derived stroma cell line MS-5. We co-cultured ALL blasts with MS-5 to develop an assay system that allows easy quantitative assessment of ALL cell growth.…”

Stromal cell-dependent ex vivo expansion of human cord blood progenitors and augmentation of transplantable stem cell activity

“…MS-5 is one such cell line, derived from noninfected Dexter-type murine long-term marrow culture, which supports CFU-S maintenance 21 and acts synergistically with human growth factors to stimulate the formation of blast colonies and macroscopic colonies from CD34 + CD38 Ϫ primitive progenitors in short-term methylcellulose assays. 22 MS-5, alone 23 or in combination with SCF and G-CSF, 24,25 also supports the proliferation of primitive lymphohematopoietic progenitors that are capable of differentiation along both myeloerythroid and lymphoid lineages and their differentiation toward the B cell lineage. Furthermore, a recent study has demonstrated that, in the presence of multiple cytokines, MS-5 cells promote a net increase of LTC-IC through self-renewal divisions.…”