High-Resolution Separation of Rare Cell Types

Leary, James F.; Ellis, Steven P.; McLaughlin, Scott R.; Corio, Mark A.; Hespelt, Steven; Gram, Janet G.; Burde, Stefan

doi:10.1021/bk-1991-0464.ch002

Cited by 16 publications

(5 citation statements)

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“…For fabricating transplantable tissues in vitro, effective cell separation methods that can provide an adequate purity, yield, and function after separation have been desired, because the purity of cells or individual cell contents in cocultured cells is important for fabricating functional tissues. , To date, the various types of cell separation methods have been developed such as field-flow fractionation (FFF), , affinity adsorption, , and flow sorting. , Especially, fluorescence-activated cell sorting (FACS) and magnetic cell sorting (MACS) are widely used as precise cell separation methods. − However, these cell separation methods require the modification of cell surfaces with fluorescent antibody or magnetic particles, leading to a serious problem upon the transplantation of separated cells to human body. Therefore, a cell separation method that requires no modification on the surface of cell is preferable for using separated cells for transplantation.…”

Section: Introductionmentioning

confidence: 99%

“…19,20 To date, the various types of cell separation methods have been developed such as field-flow fractionation (FFF), 21,22 affinity adsorption, 23,24 and flow sorting. 25,26 Especially, fluorescence-activated cell sorting (FACS) and magnetic cell sorting (MACS) are widely used as precise cell separation methods. 25−27 However, these cell separation methods require the modification of cell surfaces with fluorescent antibody or magnetic particles, leading to a serious problem upon the transplantation of separated cells to human body.…”

Section: ■ Introductionmentioning

confidence: 99%

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Hydrophobized Thermoresponsive Copolymer Brushes for Cell Separation by Multistep Temperature Change

et al. 2013

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For preparing a thermally modulated biointerface that separates cells without the modification of cell surfaces for regenerative medicine and tissue engineering, poly(N-isopropylacrylamide-co-butyl methacrylate) (P(IPAAm-co-BMA), thermo-responsive hydrophobic copolymer brushes with various BMA composition were formed on glass substrate through a surface-initiated atom transfer radical polymerization (ATRP). Characterization of the prepared surface was performed by X-ray photoelectron spectroscopy (XPS), attenuated total reflection Fourier transform infrared spectroscopy (ATR/FT-IR), and gel-permeation chromatography (GPC) measurement. Prepared copolymer brush surfaces were characterized by observing the adhesion (37 °C) and detachment (20 or 10 °C) of four types of human cells: human umbilical vein endothelial cells (HUVECs), normal human dermal fibroblasts (NHDFs), human aortic smooth muscle cells (SMCs), and human skeletal muscle myoblast cells (HSMMs). HUVECs and NHDFs exhibited their effective detachment temperature at 20 and 10 °C, respectively. Using cells' intrinsic temperature sensitivity for detachment from the copolymer brush, a mixture of green fluorescent protein (GFP)-expressing HUVECs (GFP-HUVECs) and NHDFs was separated.

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Section: Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Hydrophobized Thermoresponsive Copolymer Brushes for Cell Separation by Multistep Temperature Change

et al. 2013

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“…The analysis and/or sorting of rare cell subpopulations to a high degree of purity by flow cytometry (FCM)/cell sorting techniques has been described in a variety of biomedical applications (2)(3)(4)(6)(7)(8)11,13,15,18,22). FCM techniques can also be used to enrich samples of more commonly occurring cells.…”

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Theoretical basis for sampling statistics useful for detecting and isolating rare cells using flow cytometry and cell sorting

et al. 1997

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This paper describes new approaches to calculating the number of cells that need to be processed using flow cytometry (FCM) techniques and the subsequent time required in order to isolate a specific number of cells having selected characteristics. The methods proposed use probabilistic assumptions about the contents of the sample to be sorted, logarithmic/exponential transformations to avert the computer “underflow” and “overflow” limitations of brute force calculations for the parameters of the binomial distribution imposed by existing computer hardware, and an established mathematical procedure for calculating error bounds for the normal approximation to the binomial distribution. Estimates are derived for the total number of cells in the FCM sample volume that must be available for processing and, for given FCM cell sorting decision speeds, the total elapsed times necessary to conduct particular experiments. The proposed approach obviates the need to resort to calculation expediencies such as the theoretically limited Poisson approximation for what can be considered a Bernoulli process mathematically characterized by the binomial distribution. Tables and graphs illustrate the projected times required to complete FCM experiments as a function of “effective” cell sorting decision speeds. Results from this paper also demonstrate that, as the “effective” cell sorting decision speed increases, there may not be a corresponding linear decrease in the time required to sort a given number of cells with selected statistical properties. The focus of this paper is on the use of innovative mathematical techniques for the design of experiments involving rare cell sorting. However, these same computational approaches may also prove useful for the high‐speed enrichment sorting of non‐rare cell subpopulations. Cytometry 27:233–238, 1997. © 1997 Wiley‐Liss, Inc.

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“…Thus, individual beads from a library having high fluorescence intensities can be easily detected by flow cytometry for the ultimate sorting of aptamer beads into microcentrifuge tubes for rapid PCR based sequencing [113]. The James Leary's custom-built HiReCS flow cytometer instrumentation constitutes a high-throughput screening device that can analyze beads at rates in excess of 10 4 beads per second [114][115][116][117][118], allowing the analysis of a very small fraction of fluorescence aptamer beads to be detected and sorted from large libraries (e.g. >10 8 beads).…”

Section: Screening Of Bead Based S-odn or S 2 -Odn Librariesmentioning

confidence: 99%

Progress in Thioaptamer Development

Yang¹,

Gorenstein²

2004

CDT

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Thioaptamers are thiophosphate ester modified nucleic acids that are isolated via in vitro or bead-based thioaptamer selection against a target molecule such as a protein. Thioaptamers offer advantages over traditional aptamers in their enhanced affinity and specificity and higher stability, largely due to the properties of the sulfur backbone-modifications. An in vitro thioaptamer selection procedure that simultaneously selects for sequence and optimized hybrid phosphoromonothioate or phosphate backbone substitutions is outlined. A novel bead-based thioaptamer selection protocol that can produce mixed phosphorodithioate, phosphoromonothioate or phosphate hybrid backbones is also described. Several examples of thioaptamers targeting specific protein are provided. Such thioaptamers are shown to modulate protein activity in vivo.

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High-Resolution Separation of Rare Cell Types

Cited by 16 publications

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Hydrophobized Thermoresponsive Copolymer Brushes for Cell Separation by Multistep Temperature Change

Hydrophobized Thermoresponsive Copolymer Brushes for Cell Separation by Multistep Temperature Change

Theoretical basis for sampling statistics useful for detecting and isolating rare cells using flow cytometry and cell sorting

Progress in Thioaptamer Development

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