Improved sedimentation field-flow fractionation separation channel for concentrated cellular elution

Mélin, Carole; Lacroix, Aurélie; Lalloué, Fabrice; Pothier, A.; Zhang, L.Y.; Perraud, Aurélie; Dalmay, Claire; Lautrette, Christophe; Jauberteau, Marie‐Odile; Cardot, Philippe; Mathonnet, Muriel; Battu, Serge

doi:10.1016/j.chroma.2013.05.067

Cited by 6 publications

(7 citation statements)

References 41 publications

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“…Moreover, the use of optimum elution conditions enabling hyperlayer elution mode led to cell sorting on the basis of intrinsic cell functionality, integrity, proliferation, and maturation steps, which is essential for TICs [25,26]. In the hyperlayer elution mode, cells are driven away from the accumulation wall, avoiding harmful contact with the channel surface, and sorted with an elution order depending on cell size, 1. by means of the field and flow-rate dependence of the retention ratio R obs (R obs =tm/tr); 2. by their elution away from the channel wall with the value of s, the average distance from the center of the cell to the channel wall, greater than particle radius r; and 3. by the decrease of cell diameter between F1 and F3 fractions.…”

Section: Tic-enriched Subpopulations Obtained By Sdfffmentioning

confidence: 99%

“…Recently, we revealed the ability of SdFFF to generate enriched fractions with TIC characteristics [25,26]. SdFFF had successfully sorted colorectal TIC phenotypes on the basis of CD44 + , EpCAM high , and CD166 + expression in eight cell lines.…”

Section: Introductionmentioning

confidence: 99%

“…Magnification×25density, and rigidity; large and less dense cells are focused in the faster streamlines to be eluted first. The hyperlayer elution of HCT116 and WiDr was previously demonstrated[25,26]:…”

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confidence: 98%

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New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

et al. 2015

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Despite effective treatments, relapse of colorectal cancer (CRC) is frequent, in part caused by the existence of tumor-initiating cells (TICs). Different subtypes of TICs, quiescent and activated, coexist in tumors, defining the tumor aggressiveness and therapeutic response. These subtypes have been sorted by hyperlayer sedimentation field-flow fractionation (SdFFF) from WiDr and HCT116 cell lines. On the basis of a new strategy, including TIC SdFFF sorting, 3D Matrigel amplification, and grafting of corresponding TIC colonies on the chick chorioallantoic membrane (CAM), specific tumor matrices could be obtained. If tumors had similar architectural structure with vascularization by the host system, they had different proliferative indices in agreement with their initial quiescent or activated state. Protein analysis also revealed that tumors obtained from a population enriched for "activated" TICs lost "stemness" properties and became invasive. In contrast, tumors obtained from a population enriched for "quiescent" TICs kept their stemness properties and seemed to be less proliferative and invasive. Then, it was possible to produce different kinds of tumor which could be used as selective supports to study carcinogenesis and therapy sensitivity.

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Section: Tic-enriched Subpopulations Obtained By Sdfffmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

et al. 2015

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“…The hyperlayer elution mode was first determined based on the field and flow rate dependence of R obs . ,,, Then, at equivalent flow rates, the increase in field strength focused cells in slower stream lines, increasing retention and decreasing R obs . In that way, U87-MG showed a decrease in R obs of 33% by increasing the field from 10 to 20 g and 13% for LN18 by increasing the field from 20 to 30 g. This low difference could be explained by the fact that LN18 was eluted with higher field strength.…”

Section: Resultsmentioning

confidence: 99%

“…The advantage that SdFFF holds over other sorting techniques is the lack of need for immunolabeling. ,, It relies on cell biophysical properties: size, density, and rigidity. The cells are subjected in an empty ribbon-like separation channel (no stationary phase) to two types of forces (1) hydrodynamic lift forces generated by flowing a carrier liquid through the channel and (2) an external field applied perpendicularly to the flow direction. ,, Cells are then eluted under the “Hyperlayer” elution mode, a size/density driven separation mechanism, that ensures a drastic limitation of cell-solid panel interactions.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Glioblastoma Cancer Stem Cells Sorted by Sedimentation Field-Flow Fractionation Using an Ultrahigh-Frequency Range Dielectrophoresis Biosensor

et al. 2021

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Cancer Stem Cells (CSC) appear to be an essential target for cancer therapies, in particular in brain tumors such as Glioblastoma. Nevertheless, their isolation is made difficult by their low content in culture or tumors (< 5% of the tumor mass), and is essentially based on the use of fluorescent or magnetic labeling techniques, increasing the risk of differentiation induction. The use of label-free separation methods such as sedimentation field flow fractionation (SdFFF) is promising, but it becomes necessary to consider a coupling with a detection and characterization method for future identification and purification of CSCs from patient-derived tumors. In this study we demonstrate for the first time the capability of using an Ultra High Frequency range Dielectrophoresis (UHF-DEP) fluidic biosensor as a detector. This implies an important methodological adaptation of SdFFF cell sorting by the use of a new compatible carrier liquid DEP buffer (DEP-B). After SdFFF sorting, subpopulation derived from U87-MG and LN18 cells lines, undergo biological characterization, demonstrating that by using DEP-B as a carrier liquid, we sorted by SdFFF subpopulations with specific differentiation characteristics: F1 = differentiated cells / F2= CSCs. These sub-populations presenting high frequency crossover values similar to those measured for standard differentiated (around 110 MHz) and CSC (around 80 MHz) populations. This coupling appeared as a promising solution for the development of an online integration of these two complementary label-free separation/detection technologies.

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Field flow fractionation (FFF): practical and experimental aspects

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Battu

et al. 2022

Particle Separation Techniques

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Improved sedimentation field-flow fractionation separation channel for concentrated cellular elution

Cited by 6 publications

References 41 publications

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

Characterization of Glioblastoma Cancer Stem Cells Sorted by Sedimentation Field-Flow Fractionation Using an Ultrahigh-Frequency Range Dielectrophoresis Biosensor

Field flow fractionation (FFF): practical and experimental aspects

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