Stem cell growth factor: in situ hybridization analysis on the gene expression, molecular characterization and in vitro proliferative activity of a recombinant preparation on primitive hematopoietic progenitor cells.
Stem cell growth factor: in situ hybridization analysis on the gene expression, molecular characterization and in vitro proliferative activity of a recombinant preparation on primitive hematopoietic progenitor cells.
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DOI：
10.1038/sj.thj.6200118
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发表时间：
2001-01-01
期刊：
影响因子：
--
通讯作者：
Sugimoto, S
中科院分区：
文献类型：
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作者：
Hiraoka, A;Yano Ki, K;Sugimoto, S
导语：用干细胞生长因子（SCGF）反义探针对小鼠全胎及其胫骨原位杂交发现，SCGF mRNA在骨组织周围特异性表达，特别是在骨髓细胞、增殖软骨细胞、软骨膜和骨膜中，但在静息或肥厚软骨细胞中表达很少。方法：从scgf - α基因转染CHO细胞的条件培养基中纯化重组人（rh） scgf - α。通过内切- o-糖苷酶和唾液酸酶的酶切，rhscgf - α的分子量从45 kDa降至40 kDa，表明rhscgf - α与唾液酸发生了o-糖基化。结果：对于人骨髓CD34+Lin-细胞，rhscgf - α单独不刺激集落形成，但小簇形成（10.3 +/- 2.5/1 × 10(3) CD34+Lin-细胞）。在促红细胞生成素和GM集落刺激因子（CSF）或G-CSF的原代培养中，分别促进红细胞和粒细胞/巨噬细胞（GM）集落的生长，并在短期液体培养中进一步支持GM祖细胞。相反，rhscgf - α抑制干细胞因子（SCF）刺激的红细胞爆发，表明SCGF和SCF之间存在竞争相互作用。rhscgf - α可与白细胞介素-3和flt3配体协同促进GM集落生长，但与诱导GM祖细胞体外扩增的配体无协同作用。结论：SCGF可由骨和造血基质细胞选择性产生，并可介导其对原始造血祖细胞的增殖活性。
INTRODUCTION: In situ hybridization of whole mouse fetuses and their tibias with a stem cell growth factor (SCGF) antisense probe demonstrated specific expression of SCGF mRNA around skeletal tissues, particularly in bone marrow cells, proliferating chondrocytes, the perichondrium and periosteum, but little expression in resting or hypertrophic chondrocytes.METHODS: Recombinant human (rh) SCGF-alpha was purified from a conditioned medium of SCGF-alpha gene-transfected CHO cells. The molecular mass of rhSCGF-alpha, 45 kDa, was shifted down to 40 kDa by digestion with endo-O-glycosidase and sialidase, suggesting O-glycosylation of rhSCGF-alpha with sialic acids.RESULTS: For human bone marrow CD34+Lin- cells, rhSCGF-alpha alone did not stimulate colony-formation, but small cluster-formation (10.3 +/- 2.5/1 x 10(3) CD34+Lin- cells). It promoted growth of erythroid and granulocyte/macrophage (GM) colonies in the primary culture with erythropoietin and GM colony-stimulating factor (CSF) or G-CSF, respectively, and further supported GM progenitor cells in a short-term liquid culture. In contrast, rhSCGF-alpha suppressed stem cell factor (SCF)-stimulated erythroid bursts, indicating some competitive interaction between SCGF and SCF. rhSCGF-alpha was synergistic with interleukin-3 and the flt3 ligand to enhance GM colony-growth, but not synergistic with those inducing ex vivo expansion of GM progenitor cells.CONCLUSION: SCGF is selectively produced by osseous and hematopoietic stromal cells, and can mediate their proliferative activity on primitive hematopoietic progenitor cells.