Carboxyfluorescein and biotin neuromedin C analogues: Synthesis and applications

Walker, Brian; Gray, J.; Burns, Deirdre; Wang, Qiming J.; Adrian, Thomas E.; Nichols, David H.; Murphy, Richard F.; Nelson, Jane

doi:10.1016/0196-9781(94)00174-x

Cited by 4 publications

(3 citation statements)

References 25 publications

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“…26 CFSE was also used to track the fate of human intervertebral disc cells in vitro. 23 Localization of CFSE-labeled cells is useful since the dye does not passively transfer to unlabeled cells 39 and has been shown to not transfer between labeled and unlabeled cells in coculture experiments in vitro. 36 CFSE labeling may be applicable in cartilage repair studies in vivo since S and M zone chondrocytes isolated from cartilage of 4-6-month-old Yucatan mini-pigs have also been effectively labeled, 21 and could be used in conjunction with PKH26, another fluorescent dye previously described for tissue engineering applications.…”

Section: Figmentioning

confidence: 99%

Tracking Chondrocytes and Assessing Their Proliferation with Carboxyfluorescein Diacetate Succinimidyl Ester: Effects on Cell Functions

Chawla

Masuda

Sah

2010

Tissue Engineering Part C: Methods

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Distinguishing between implanted and host-derived cells, as well as cells of different phenotypes, is important in determining mechanisms of cell-based repair of cartilage. The objectives of this study were to assess the utility of carboxyfluorescein diacetate, succinimidyl ester (''CFDA, SE'' or CFSE) for tracking chondrocytes from superficial (S) and middle (M) zones and their proliferation, and to determine the effects of CFSE on the chondrocyte functions, proliferation, and synthesis of proteoglycan 4 (PRG4) and glycosaminoglycan (GAG). CFSE-labeled and unlabeled S and M zone chondrocytes were plated in either low-or high-density (10,000 or 200,000 cells=cm 2 ) monolayer, incubated, and analyzed on days 1 and 7. Cell suspensions were analyzed for retention of CFSE by flow cytometry and fluorescence microscopy and for cell proliferation by assay for DNA and GAG. Cultures were also analyzed for newly synthesized PRG4. Deconvolution of flow cytometric histograms was done to determine the number of cells in each doubling generation. Most chondrocytes were labeled consistently and intensely labeled with CFSE through 10 cycles of division. At day 7 of culture, approximately 95% of S and M zone cells seeded at high density could be distinguished as fluorescent. Chondrocyte proliferation and synthesis of PRG4 were unaffected by cell labeling, while GAG synthesis was slightly diminished. CFSE may be useful in determining the fate and function of implanted chondrocytes in vivo.

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Section: Figmentioning

confidence: 99%

Tracking Chondrocytes and Assessing Their Proliferation with Carboxyfluorescein Diacetate Succinimidyl Ester: Effects on Cell Functions

Chawla

Masuda

Sah

2010

Tissue Engineering Part C: Methods

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“…This assay utilized a rabbit GRP 18-27 antiserum raised by this group and described previously (Walker et al, 1995). Five thousand c.p.m.…”

Section: Radioimmunoassaymentioning

confidence: 99%

Breast cancer cell-associated endopeptidase EC 24.11 modulates proliferative response to bombesin

Burns

Walker²,

Gray³

et al. 1998

Br J Cancer

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SummaryWe have investigated the production, growth and inactivation of gastrin-releasing peptide (GRP)-like peptides in human breast cancer cell lines. Radioimmunoassay detected GRP-like immunoreactivity (GRP-LI) in T47D breast cancer cells but not in the conditioned medium, indicating rapid clearance. No GRP-LI was found in the ZR-75-1 or MDA-MB-436 cells or their conditioned medium. Highperformance liquid chromatography (HPLC) analysis of the GRP-LI in the T47D cells revealed a major peak, which co-eluted with GRP 18-27 , and a minor more hydrophilic peak. In vitro stimulation of T47D cell growth by bombesin (BN) was enhanced to 138% of control levels (bombesin alone) by the addition of the selective endopeptidase EC 3.4.24.11 inhibitor phosphoramidon (0.1 ng ml -1 ). Fluorogenic analysis using whole cells confirmed low levels of this phosphoramidon-sensitive enzyme on the T47D cells. This enzyme, previously unreported in human breast cancer cells, significantly modulates both T47D growth and its response to BN-induced growth.

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“…Clearly, eosinophils are involved in immunological function. Beyond that, Walker et al (1995) reported the binding of carboxyfluorescein NC to a rat stomach cell population tentatively identified as eosinophils. This raised the possibility that eosinophils interact with peptides related not only to immune function, but to gut neural and secretory function as well.…”

mentioning

confidence: 99%

Peptides bind to eosinophils in the rat stomach

Nichols¹,

Lovas²,

Adrian³

et al. 1998

Anat. Rec.

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Carboxyfluorescein and biotin neuromedin C analogues: Synthesis and applications

Cited by 4 publications

References 25 publications

Tracking Chondrocytes and Assessing Their Proliferation with Carboxyfluorescein Diacetate Succinimidyl Ester: Effects on Cell Functions

Tracking Chondrocytes and Assessing Their Proliferation with Carboxyfluorescein Diacetate Succinimidyl Ester: Effects on Cell Functions

Breast cancer cell-associated endopeptidase EC 24.11 modulates proliferative response to bombesin

Peptides bind to eosinophils in the rat stomach

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