Basic Fibroblast Growth Factor Stimulates the Competence Phase of Subcultured Endothelial Cells and the Progression Phase of Primary Cultured Smooth Muscle Cells from Rat Aorta.

Nagaura, Takeshi; Okabe, Motonori; Kobayashi, Shogo; Kimura, Ikuko; Kimura, Masayasu

doi:10.1248/bpb.17.1176

Cited by 9 publications

(6 citation statements)

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“…bFGF is also a potent mitogen for EC proliferation since the bFGF effect is enhanced in an autocline manner in vitro (8). In the previous study, we demonstrated that bFGF hastens the starting time of [3H]-thymidine incor poration into subcultured EC of rat aorta, suggesting the competence effect of bFGF (9). However, it remains to be determined if bFGF is a competence factor in EC as well as in fibroblasts.…”

mentioning

confidence: 83%

Competence Effect of Basic Fibroblast Growth Factor on Cell Cycle in Subcultured Endothelial Cells of Rat Aorta by Flow Cytometry

Kimura

Okabe

Ogasawara

et al. 1996

Japanese Journal of Pharmacology

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ABSTRACT-With flow cytometry, we investigated the effect of basic fibroblast growth factor (bFGF)-induced competence in subcultured endothelial cells (EC) (4-9-passage) of rat thoracic aorta. The cell population in each phase of the cell cycle was determined by a double staining technique with fluorescein isothiocyanate-conjugated mouse monoclonal antibody against the proliferation-associated nucleus antigen Ki-67 and propidium iodide for total DNA content. EC were cultured in medium containing 5% fetal bo vine serum (FBS) for 6 days. After serum-starvation for 2 days, the treatment with bFGF (3-10 ng/ml) for 12 hr promoted the entry of cells into the G1 phase from the G0 phase concentration-dependently.bFGF (10 ng/ml) increased the cell population in the G1 phase by 50% of the total EC, compared with the control culture without bFGF. A further 12-15-hr culture with 1% FBS after bFGF treatment promoted the entry of the cell into the S phase. Thus flow cytometric analysis demonstrates that bFGF stimulates the entry of EC into the G1 phase from the Go phase. Keywords:Basic fibroblast growth factor, Flow cytometry, Rat aortic endothelial cell, Ki-67 antigen, Propidium iodideAbnormal proliferation of vascular endothelial cells (EC) causes many diseases such as diabetic retinopathy (1) and atherosclerosis (2). Development of these angio genic diseases are reported to be induced several growth factors such as basic fibroblast growth factor (bFGF), platelet-derived growth factor and interleukin-8 (3 5). Growth factors are classified into two types, a compe tence factor and a progression factor, based on their modes of enhancement of cell proliferation (6). The com petence factors advance the cell cycle from the Go to G1 phase, whereas the progression factors promote entrance to the S phase from the G1 phase (7). bFGF, a compe tence factor not a progression factor, can induce DNA synthesis only in combination with a progression factor containing platelet-poor plasma in Balb/3T3 fibroblast (7). bFGF is also a potent mitogen for EC proliferation since the bFGF effect is enhanced in an autocline manner in vitro (8). In the previous study, we demonstrated that bFGF hastens the starting time of [3H]-thymidine incor poration into subcultured EC of rat aorta, suggesting the competence effect of bFGF (9). However, it remains to be determined if bFGF is a competence factor in EC as well as in fibroblasts.A proliferation-associated nucleus antigen, Ki-67, is a useful tool for detecting the transition from the Go to G1 phase, because this antigen is expressed in actively grow ing cells but not in Go-arrested (quiescent) cells (10, 11). In the present study using flow cytometry with double staining of Ki-67 antigen and total DNA, we provided evidence that bFGF stimulates the competence in quies cent EC of rat thoracic aorta. MATERIALS AND METHODS Cell cultureRat aortic EC in primary culture were prepared by the method reported by Schwartz (12) and Nagaura et al. (9) as follows: The thoracic aorta of male Wistar rats (9 week-old, 24...

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confidence: 83%

Competence Effect of Basic Fibroblast Growth Factor on Cell Cycle in Subcultured Endothelial Cells of Rat Aorta by Flow Cytometry

Kimura

Okabe

Ogasawara

et al. 1996

Japanese Journal of Pharmacology

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“…Both acidic and basic FGF induce endothelial cell proliferation and play a major role in the proliferative phase of wound repair, but a minor role in the preinflammatory and inflammatory phases. aFGF is mitogenic for dermal fibroblasts and epidermal keratinocytes and bFGF stimulates the proliferation of fibroblasts and vascular smooth muscle cells 11 . In the normal wound healing process there is a lag before the proliferative phase begins while components and factors from the preliminary and inflammatory phases move into the wound site.…”

Section: Recent Research and Its Implications For Clinical Wound Healingmentioning

confidence: 99%

“…aFGF is mitogenic for dermal fibroblasts and epidermal keratinocytes and bFGF stimulates the proliferation of fibroblasts and vascular smooth muscle cells. 11 In the normal wound healing process there is a lag before the proliferative phase begins while components and factors from the preliminary and inflammatory phases move into the wound site.…”

Section: Recent Research and Its Implications For Clinical Wound Healingmentioning

confidence: 99%

Increased healing of the disrupted wound—A myth or simply enhanced angiogenesis following wounding? Personal discussions with Tom Hunt from the 1960s to the present

Cherry

Hughes

2003

Wound Repair Regeneration

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“…Rat aortic EC in primary culture were prepared as shown previously (5,6 with an optical microscope. The isolated EC were treated with Ca 2+, Mg2+-free phosphate-buffered saline (PBS) containing 0.5 mg/ml trypsin and 0.54 mM EDTA and subcultured at a split ratio of 1 : 4.…”

Section: Cell Culturementioning

confidence: 99%

“…The subcultured EC between the 4th and 9th passage were used in the experiments. EC were identified with their typical cobble stone-like morphology (6) and the ability to take up acetylated-low density lipoprotein labeled with the fluores cent probe 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbo cyanine perchlorate (Dil-Ac-LDL; Biomedical Technol ogies, Stoughton, MA, USA) (7). Cells were incubated with 10 ,ug/ml Dil-Ac-LDL at 371C in DMEM for 4 hr.…”

Section: Cell Culturementioning

confidence: 99%

Platelet-Derived Growth Factor Blocks the Cell-Cycle Transition from the G0 to G1 Phase in Subcultured Angiogenic Endothelial Cells in Rat Thoracic Aorta.

et al. 1997

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ABSTRACT-Platelet-derived growth factor (PDGF)-BB induces tube formation by the differentiating (tube-forming) endothelial cells (EC) of rat thoracic aorta, although PDGF-BB does not affect the proliferative EC (increasing the cell numbers) at the progression phase. These changes in the responses to PDGF-BB were due to the phenotype-dependent expression of PDGF p-receptor (PDGFR-j3) on EC be cause PDGFR-p-like immunoreactivity was observed in the angiogenic EC forming a tube-like structure in 35-day culture with 10% fetal bovine serum, but not in the proliferative EC in 5-day culture. To elucidate the functional role of PDGFR-j3 in the alteration of EC phenotype, the influence of PDGF-BB on the cell cycle of EC was investigated by flow cytometry. This analysis demonstrates that PDGF-BB blocks the transition from the Go to G1 phase in the 35-day cultured EC, although no effect was observed on any phases of the cell cycle in 5-day culture. We conclude that 1) PDGFR-j3 is expressed in mature angiogenic EC of rat aorta, and 2) PDGF-BB may contribute to promotion of the EC differentiation with tubular morphogenesis by inhibiting cell growth.

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Basic Fibroblast Growth Factor Stimulates the Competence Phase of Subcultured Endothelial Cells and the Progression Phase of Primary Cultured Smooth Muscle Cells from Rat Aorta.

Cited by 9 publications

References 0 publications

Competence Effect of Basic Fibroblast Growth Factor on Cell Cycle in Subcultured Endothelial Cells of Rat Aorta by Flow Cytometry

Competence Effect of Basic Fibroblast Growth Factor on Cell Cycle in Subcultured Endothelial Cells of Rat Aorta by Flow Cytometry

Increased healing of the disrupted wound—A myth or simply enhanced angiogenesis following wounding? Personal discussions with Tom Hunt from the 1960s to the present

Platelet-Derived Growth Factor Blocks the Cell-Cycle Transition from the G0 to G1 Phase in Subcultured Angiogenic Endothelial Cells in Rat Thoracic Aorta.

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