Pre-apoptotic sub-population cell sorting from diosgenin apoptosis induced 1547 cells by Sedimentation Field-Flow Fractionation

Bégaud-Grimaud, G.; Battu, Serge; Liagre, Bertrand; Léger, David Y.; Beneytout, Jean-Louis; Cardot, Philippe

doi:10.1016/j.chroma.2006.06.065

Cited by 26 publications

(61 citation statements)

References 44 publications

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“…As previously described (22,27,33,39,44,45), cell species are eluted under the Hyperlayer model, a size/density dependent cell elution mode (31,(33)(34)(35)(36)(37)(38)(39), which predicts that large and weaker cells are focused in faster streamlines to be eluted first. Hyperlayer elution mode allowed, by limiting cell-accumulation wall interactions, a cell elution which: i) respected repeatability, reproducibility and recovery; ii) respected cell functional integrity; iii) provided high levels of short and long-term viability (no apoptosis induction); iv) respected the maturation and differentiation stages; and finally; v) enhanced better sub-population sorting and characterization (20,22,26,27).…”

Section: Discussionmentioning

confidence: 99%

“…In order to separate sensitive and resistant cells to Fas-induced apoptosis, we have studied the apoptotic response in different fractions obtained after SdFFF U-87 MG cell sorting, a non-invasive and tagless method for cell separation that was previously used in different application fields of oncology (27,32,33,(41)(42)(43). Different aspects have been evaluated including: i) monitoring of the apoptosis or differentiation induction (32,33,42,43); ii) cell sorting of specific subpopulations [pre-apoptotic (33), differentiated cells (32)]; iii) kinetics of apoptosis (27). SdFFF was also used to sort specific phenotypes from complex cancer cell populations such as neuroblastomas (26), or to sort immature cancer cells (22), two applications which can be directly used in this study.…”

Section: Discussionmentioning

confidence: 99%

“…R obs F3 = 0.351±0.021 (n>6). As previously described (32,33), the absolute R obs value particularly depends on culture conditions, thus for U-87 MG cells the number of cell passages which should range from 2 to 8. At constant flow rate (0.80 ml/min), according to the 'Hyperlayer' elution mode description (31,(33)(34)(35)(36)(37)(38)(39), the R obs value for F3 is field dependant: R obs = 0.366±0.012 (n=3) at 20.00 g. By using the following equation (40):…”

Section: Distinct Cell Populations Identified By Sdfff Elutionmentioning

confidence: 99%

“…Two methods were used to obtain immature and tumor cell stems, a cell-sorting method, the sedimentation field flow fractionation (SdFFF) able to separate immature and differentiated cells as previously demonstrated (20)(21)(22) and a longest selection method based on the growth properties of tumor stem cells in a defined culture medium (23).…”

Section: Introductionmentioning

confidence: 99%

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Cancer stem cells from human glioma cell line are resistant to Fas-induced apoptosis

Jauberteau¹

2009

Int J Oncol

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Abstract. Glioblastoma is the most common primary brain tumor, characterized by its resistance to treatments. To define efficient therapy, the origin of tumor-forming cells needs to be elucidated in order to search for new therapeutic pathways. The objective of this study was to determine the different cell populations constituting a human glioblastoma cell line, U-87 MG and their sensitivity to apoptosis induced through the activation of Fas, a membranous death receptor. By a cell sorting method, the sedimentation field flow fractionation, two major cell subpopulations were identified, a most differentiated cell fraction, containing large and adherent cells, sensitive to Fas-induced apoptosis and another one, characterized by small cells forming aggregates, expressing CD133, a marker of stem cells and more resistant to Fas-activated apoptosis. By using a selective method of culture, adapted for neural stem cell cultures, we have verified that the U-87 MG cell line contained cancer stem cells similar to the immature ones obtained by the cell sorting method. Interestingly, while these tumor stem cells, expressing CD133, were resistant to Fas-induced apoptosis, monomeric form of Fas protein was detected predominantly in these cells. In contrast, the most mature cells, responsive to Fas-activated apoptosis, collected in another cell fraction, contained oligomeric aggregates of Fas protein, a pre-signalling form of the Fas receptor, essential for the initiation of apoptosis through its activation. These results suggest that these immature stem cells in glioma could be an important factor of resistance to chemotherapy requiring apoptosis through Fas signalling system. Indeed, future strategies of treatment, inducing differentiation of these stem cells need to be considered to enhance therapeutic efficiency.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Distinct Cell Populations Identified By Sdfff Elutionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cancer stem cells from human glioma cell line are resistant to Fas-induced apoptosis

Jauberteau¹

2009

Int J Oncol

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“…Diosgenin (40 μM) was used because our previous studies on different cancer cell lines had shown that this concentration induced apoptosis (5)(6)(7)25,26).…”

Section: Discussionmentioning

confidence: 99%

Cyclooxygenase-2 and 5-lipoxygenase pathways in diosgenin-induced apoptosis in HT-29 and HCT-116 colon cancer cells

Lepage

Liagre²,

Cook-Moreau³

et al. 2010

Int J Oncol

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Abstract. We studied the relationship between diosgenininduced apoptosis and arachidonic acid metabolism in two cancer cell lines, the HT-29 and HCT-116. Diosgenin is a steroidal saponin known for its antiproliferative and proapoptotic effects on cancer cells. We focused our attention on two enzymes intervening in two different pathways of arachidonic metabolism: cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX). HT-29 and HCT-116 cells express 5-LOX but only HT-29 cells express COX-2 since HCT-116 are deficient. After 40 μM diosgenin treatment, we observed apoptosis hallmarks in both cell lines but HT-29 cells were more resistant with delayed apoptosis. In these cells, COX-2 expression and activity were increased and for the first time we showed that diosgenin also increased 5-LOX expression and enhanced leukotriene B 4 production. Inhibition of 5-LOX by AA-861 significantly reduced apoptosis in both cell lines but COX-2 inhibition by NS-398 strongly sensitized HT-29 cells to diosgenin-induced apoptosis compared to HCT-116 cells. In this study, we showed the implication of COX-2 and 5-LOX in diosgenin-induced apoptosis but these results demonstrate how difficult it is to assess the correlation between the apoptotic signalling pathway of diosgenin and arachidonic acid metabolism with certitude.

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Mammalian Cell Sorting with Sedimentation Field-Flow Fractionation

Bégaud-Grimaud

Battu

Léger

et al. 2011

Field-Flow Fractionation in Biopolymer Analysis

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Pre-apoptotic sub-population cell sorting from diosgenin apoptosis induced 1547 cells by Sedimentation Field-Flow Fractionation

Cited by 26 publications

References 44 publications

Cancer stem cells from human glioma cell line are resistant to Fas-induced apoptosis

Cancer stem cells from human glioma cell line are resistant to Fas-induced apoptosis

Cyclooxygenase-2 and 5-lipoxygenase pathways in diosgenin-induced apoptosis in HT-29 and HCT-116 colon cancer cells

Mammalian Cell Sorting with Sedimentation Field-Flow Fractionation

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