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经过基因工程改造以表达成纤维细胞生长因子受体1的FDC-P1髓样细胞,在成纤维细胞生长因子和肝素存在的情况下会增殖并分化。

FDC-P1 myeloid cells engineered to express fibroblast growth factor receptor 1 proliferate and differentiate in the presence of fibroblast growth factor and heparin.

作者信息

Li M, Bernard O

机构信息

Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.

出版信息

Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3315-9. doi: 10.1073/pnas.89.8.3315.

Abstract

Full-length murine fibroblast growth factor (FGF) receptor 1 (FGFR-1L) cDNA was introduced into the FDC-P1 mouse myeloid progenitor cell line, which lacks FGF receptors and depends on interleukin 3 (IL-3) or granulocyte/macrophage colony-stimulating factor (GM-CSF) for its proliferation and survival. The expression of the FGFR-1L gene in FDC-P1 cells allowed these cells to grow in the presence of FGF and heparin. The resulting cell line, designated FD FGFR-1L.A, exhibited a more mature myeloid phenotype than did the parental FD FGFR-1L cells or uninfected FDC-P1 cells. They formed mainly dispersed colonies in soft-agar cultures when grown in the presence of FGF and heparin, suggestive of myeloid differentiation. The cells can be switched between growth on FGF/heparin and IL-3. Northern blot analysis and cytochemical staining demonstrated that FD FGFR-1L.A cells expressed myeloperoxidase mRNA and protein, biochemical markers specifically expressed during differentiation from the promyelocytic to the granulocytic stages, whereas the parental FD FGFR-1L cells and FDC-P1 cells failed to express this marker. These results indicate that the expression of FGFR-1L by FDC-P1 cells transmitted signals for growth in the presence of FGF and heparin and generated an additional signal for early myeloid differentiation but failed to commit FD FGFR-1L.A cells to terminal differentiation. This in vitro culture system can be used for molecular analysis of the regulation of cellular growth and differentiation mediated by the FGFs and their receptors.

摘要

将全长小鼠成纤维细胞生长因子(FGF)受体1(FGFR - 1L)的互补DNA(cDNA)导入FDC - P1小鼠骨髓祖细胞系,该细胞系缺乏FGF受体,其增殖和存活依赖于白细胞介素3(IL - 3)或粒细胞/巨噬细胞集落刺激因子(GM - CSF)。FGFR - 1L基因在FDC - P1细胞中的表达使这些细胞能够在FGF和肝素存在的情况下生长。由此产生的细胞系命名为FD FGFR - 1L.A,与亲代FD FGFR - 1L细胞或未感染的FDC - P1细胞相比表现出更成熟的骨髓表型。当在FGF和肝素存在的情况下生长时,它们在软琼脂培养物中主要形成分散的集落,提示骨髓分化。这些细胞可以在FGF/肝素和IL - 3上生长之间切换。Northern印迹分析和细胞化学染色表明,FD FGFR - 1L.A细胞表达髓过氧化物酶mRNA和蛋白,这是从早幼粒细胞到粒细胞阶段分化过程中特异性表达的生化标志物,而亲代FD FGFR - 1L细胞和FDC - P1细胞未能表达该标志物。这些结果表明,FDC - P1细胞中FGFR - 1L的表达在FGF和肝素存在的情况下传递生长信号,并产生早期骨髓分化的额外信号,但未能使FD FGFR - 1L.A细胞进行终末分化。这种体外培养系统可用于对由FGF及其受体介导的细胞生长和分化调控进行分子分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/effe/48857/4f3dec2cc1de/pnas01082-0162-a.jpg

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