Sedimentation field flow fractionation purification of immature neural cells from a human tumor neuroblastoma cell line

Lautrette, Christophe; Cardot, Philippe; Vermot-Desroches, Claudine; Wijdenes, John; Jauberteau, Marie‐Odile; Battu, Serge

doi:10.1016/s1570-0232(03)00229-0

Cited by 32 publications

(53 citation statements)

References 43 publications

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“…4), leading, as for neuroblastoma cell line (26), to complex fractograms (Fig. 3) in contrast to many other cell lines (22,27,42,43). According to results concerning: i) the particle elution away from the accumulation wall (s>r); ii) field dependence of the Robs value; and iii) the limited interactions between cells and accumulation wall leading to low reversible cellular release (RP , Fig.…”

Section: Discussionmentioning

confidence: 98%

“…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%

“…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%

“…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%

“…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: 98%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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

Jauberteau¹

2009

Int J Oncol

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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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Biocompatible channels for field-flow fractionation of biological samples: correlation between surface composition and operating performance

et al. 2005

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Biocompatible methods capable of rapid purification and fractionation of analytes from complex natural matrices are increasingly in demand, particularly at the forefront of biotechnological applications. Field-flow fractionation is a separation technique suitable for nano-sized and micro-sized analytes among which bioanalytes are an important family. The objective of this preliminary study is to start a more general approach to field-flow fractionation for bio-samples by investigation of the correlation between channel surface composition and biosample adhesion. For the first time we report on the use of X-ray photoelectron spectroscopy (XPS) to study the surface properties of channels of known performance. By XPS, a polar hydrophobic environment was found on PVC material commonly used as accumulation wall in gravitational field-flow fractionation (GrFFF), which explains the low recovery obtained when GrFFF was used to fractionate a biological sample such as Staphylococcus aureus. An increase in separation performance was obtained first by conditioning the accumulation wall with bovine serum albumin and then by using the ion-beam sputtering technique to cover the GrFFF channel surface with a controlled inert film. XPS analysis was also employed to determine the composition of membranes used in hollow-fiber flow field-flow fractionation (HF FlFFF). The results obtained revealed homogeneous composition along the HF FlFFF channel both before and after its use for fractionation of an intact protein such as ferritin.

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Sedimentation field flow fractionation purification of immature neural cells from a human tumor neuroblastoma cell line

Cited by 32 publications

References 43 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

Mammalian Cell Sorting with Sedimentation Field-Flow Fractionation

Biocompatible channels for field-flow fractionation of biological samples: correlation between surface composition and operating performance

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