Biophysical Phenotyping and Modulation of ALDH+ Inflammatory Breast Cancer Stem‐Like Cells

Cancer stem‐like cells (CSCs) have been shown to initiate tumorigenesis and cancer metastasis in many cancer types. Although identification of CSCs through specific marker expression helps define the CSC compartment, it does not directly provide information on how or why this cancer cell subpopulation is more metastatic or tumorigenic. In this study, the functional and biophysical characteristics of aggressive and lethal inflammatory breast cancer (IBC) CSCs at the single‐cell level are comprehensively profiled using multiple microengineered tools. Distinct functional (cell migration, growth, adhesion, invasion and self‐renewal) and biophysical (cell deformability, adhesion strength and contractility) properties of ALDH+ SUM149 IBC CSCs are found as compared to their ALDH? non‐CSC counterpart, providing biophysical insights into why CSCs has an enhanced propensity to metastasize. It is further shown that the cellular biophysical phenotype can predict and determine IBC cells' tumorigenic ability. SUM149 and SUM159 IBC cells selected and modulated through biophysical attributes—adhesion and stiffness—show characteristics of CSCs in vitro and enhance tumorigenicity in in vivo murine models of primary tumor growth. Overall, the multiparametric cellular biophysical phenotyping and modulation of IBC CSCs yields a new understanding of IBC's metastatic properties and how they might develop and be targeted for therapeutic interventions.     

Screening Therapeutic Agents Specific to Breast Cancer Stem Cells Using a Microfluidic Single‐Cell Clone‐Forming Inhibition Assay

Screens of cancer stem cells (CSCs)‐specific agents present significant challenges to conventional cell assays due to the difficulty in preparing CSCs ready for drug testing. To overcome this limitation, developed is a microfluidic single‐cell assay for screening breast cancer stem cell–specific agents. This assay takes advantage of the single‐cell clone‐forming capability of CSCs, which can be specifically inhibited by CSC‐targeting agents. The single‐cell assay is performed on a microfluidic chip with an array of 3840 cell‐capturing units; the single‐cell arrays are easily formed by flowing a cell suspension into the microchip. Achieved is a single cell‐capture rate of ≈60% thus allowing more than 2000 single cells to be analyzed in a single test. Over long‐term suspension culture, only a minority of cells survive and form tumorspheres. The clone‐formation rate of MCF‐7, MDA‐MB‐231, and T47D cells is 1.67%, 5.78%, and 5.24%, respectively. The clone‐forming inhibition assay is conducted by exposing the single‐cell arrays to a set of anticancer agents. The CSC‐targeting agents show complete inhibition of single‐cell clone formation while the nontargeting ones show incomplete inhibition effects. The resulting microfluidic single‐cell assay with the potential to screen CSC‐specific agents with high efficiency provides new tools for individualized tumor therapy.     

Tracking and Finding Slow‐Proliferating/Quiescent Cancer Stem Cells with Fluorescent Nanodiamonds

Quiescent cancer stem cells (CSCs) have long been considered to be a source of tumor initiation. However, identification and isolation of these cells have been hampered by the fact that commonly used fluorescent markers are not sufficiently stable, both chemically and photophysically, to allow tracking over an extended period of time. Here, it is shown that fluorescent nanodiamonds (FNDs) are well suited for this application. Genotoxicity tests of FNDs with comet and micronucleus assays for human fibroblasts and breast cancer cells indicate that the nanoparticles neither cause DNA damage nor impair cell growth. Using AS‐B145‐1R breast cancer cells as the model cell line for CSC, it is found that the FND labeling outperforms 5‐ethynyl‐2′‐deoxyuridine (EdU) and carboxyfluorescein diacetate succinimidyl ester (CFSE) in regards to its long‐term tracking capability (>20 d). Moreover, through a quantification of their stem cell activity by measuring mammosphere‐forming efficiencies (MFEs) and self‐renewal rates, the FND‐positive cells are identified to have an MFE twice as high as that of the FND‐negative cells isolated from the same dissociated mammospheres. Thus, the nanoparticle‐based labeling technique provides an effective new tool for tracking and finding slow‐proliferating/quiescent CSCs in cancer research.     

Mouse embryonic stem cell sorting for the generation of transgenic mice by sedimentation field-flow fractionation

Mouse embryonic stem (ES) cells are an important tool for generation of transgenic mice and genetically modified mice. A rapid and efficient separation of ES cells that respects cell integrity, viability, and their developmental potential while also allowing purified ES fraction collection under sterile conditions might be of great interest to facilitate the generation of chimeric animals. In this study, we demonstrated for the first time the effectiveness of a sedimentation field-flow fractionation (SdFFF) cell sorter to provide, with a characteristic DNA content, a purified ES cell fraction and with a high in vivo developmental potential to prepare transgenic mice by generation of chimeras having a high percentage of chimerism.     

Effects of estimation errors on cause-selecting charts

The Cause-Selecting Chart (CSC) is a useful statistical process control tool for analyzing multistage processes. It distinguishes between incoming and outgoing quality problems by establishing a relationship between input and output measurements. In practice, the model relating the input and output must first be estimated before the CSC is implemented. Most previous work on CSCs has focused on the case when the model is estimated without error. Far less is known about the performance of CSCs with parameter estimation errors. This paper investigates the effect of parameter estimation errors on the performance of CSCs. The results indicate that the charted statistics are correlated after parameter estimation. It is also shown that this correlation will increase as the mean shift in the input increases. The implications for the use of CSCs when parameters are estimated are discussed, including the use of widened control limits and self-starting procedures.     

Biomimetic Nanobomb for Synergistic Therapy with Inhibition of Cancer Stem Cells

Cancer stem cells (CSCs), a type of cell with self-renewal, unlimited proliferation, and insensitivity to common physical and chemical factors, are the key to cancer metastasis, recurrence, and chemo-resistance. Available CSCs inhibition strategies are mainly based on small molecule drugs, yet are limited by their off-target toxicity. The link between CSCs and non-CSCs interconversion is difficult to sever. In this work, a nanotherapeutic strategy based on MnO_x-loaded polydopamine (MnO_x/PDA) nanobombs with chemodynamic, photodynamic, photothermal and biodegradation properties to inhibit CSCs and non-CSCs concurrently is reported. The MnO_x/PDA nanobombs can directly disrupt the microenvironment and tumorigenic capacity of CSCs by generating hyperthermia, oxidative stress and alleviating hypoxia. The markers of CSCs are subsequently downregulated, leading to the clearance of CSCs. Meanwhile, the synergistic therapy mediated by MnO_x/PDA nanobombs can directly ablate the bulk tumor cells, thus cutting off the supply of CSCs transformation. For tumor targeting, MnO_x/PDA is coated with macrophage membrane. The final tumor inhibition rate of the synergistic therapy is 70.8% in colorectal cancer (CRC) model. Taken together, the present work may open up the exploration of nanomaterial-based synergistic therapy for the simultaneous elimination of therapeutically resistant CSCs and non-CSCs.     

CFRP加固火灾后混凝土短柱抗震性能的试验研究

进行了1根未受火混凝土短柱、1根火灾后混凝土短柱、1根CFRP加固未受火混凝土短柱和3根CFRP加固火灾后混凝土短柱的拟静力试验,考察了CFRP的加固量和加固方式对火灾后混凝土短柱抗震加固效果的影响情况,建议了CFRP加固火灾后混凝土柱抗剪承载力的实用计算方法。研究结果表明:外包CFRP加固可将未受火混凝土短柱的抗剪承载力提高21.8%、可将火灾后混凝土短柱的抗剪承载力提升至未受火试件的111.2%~117.1%、可提高受火前后短柱的极限变形能力和累积滞回耗能。与未受火试件相比,外包CFRP不会提高火灾后加固混凝土短柱的割线刚度。对于相同的加固量,全包CFRP柱的抗剪承载力略优于条带包裹柱。受火前、后和外包CFRP加固前、后混凝土短柱的滞回规则均类似。     

Properties of anti-washout-type calcium silicate bone cements containing gelatin

Novel washout-resistant bone substitute materials consisting of gelatin-containing calcium silicate cements (CSCs) were developed. The washout resistance, setting time, diametral tensile strength (DTS), morphology, and phase composition of the hybrid cements were evaluated. The results indicated that the dominant phase of β-Ca₂SiO₄ for the SiO₂–CaO powders increased with an increase in the CaO content of the sols. After mixing with water, the setting times of the CSCs ranged from 10 to 29 min, increasing with a decrease in the amount of CaO in the sols. Addition of gelatin into the CSC significantly prolonged (P < 0.05) the setting time by about 2 and 8 times, respectively, for 5% and 10% gelatin. However, the presence of gelatin appreciably improved the anti-washout and brittle properties of the cements without adversely affecting mechanical strength. It was concluded that 5% gelatin-containing CSC may be useful as bioactive bone repair materials.     

Size determination of diesel soot particles using flow and sedimentation field-flow fractionation

The applicability of field-flow fractionation (FFF) was investigated for determination of size and size distribution of diesel soot particles. A sample preparation procedure was developed for FFF analysis where soot particles are recovered from filters in an ethanol bath sonicator, and then they are dispersed in water containing 0.05% Triton X-100 and 0.02% NaN(3). Mean diameters obtained from sedimentation FFF (SdFFF) and flow FFF (FlFFF) agree well with each other and are in good agreement with diameters obtained from photon correlation spectroscopy (PCS) and scanning electron microscopy. The relative error was less than 11%. Data show diesel soot particles have broad size distributions ranging from 0.05 up to ～0.5 μm with the mean diameters between 0.1 and 0.2 μm. The use of FlFFF is more convenient as FlFFF fractograms can be converted directly to size distributions, while the conversion of the SdFFF fractogram needs the particle density information. The density needed for SdFFF analysis was obtained by combining the SdFFF retention data with the PCS size data. For samples whose density is known, SdFFF may be more useful as SdFFF provides a wider dynamic range than FlFFF under constant field strength.     

离心沉降场流分离技术用于纳米颗粒尺寸的准确表征

以名义粒径为240nm的聚苯乙烯标准纳米颗粒悬浮液为样品,利用离心沉降场流分离仪以0．1％FL-70的水溶液为流动相分离。测定检测器波长为254nm时,纳米颗粒在离心力场的作用下通过分离通道的保留时间,从而确定纳米颗粒的平均粒径,测定值为247nm。此外,还利用扫描电镜法测定了纳米颗粒标准物质的粒径,测定值为220nm,结果表明,离心沉降场流分离技术较扫描电镜法更接近于聚苯乙烯纳米颗粒标准的名义粒径,具有更高的准确度和精密度。该方法可望成为纳米尺度表征的一种参考方法。     

Expression profile of T cell associated molecules in the interfacial tissue of aseptically loosened prosthetic joints

The involvement of T cells in the progression of inflammation in response to wear debris at the interface of aseptically loosened joints is currently undefined. This cell type has repeatedly been demonstrated to be a common component of the cellular membrane, the interface, which forms between the bone and implant of total joint replacements (TJRs) [1, 2]. Three further insights into the role of this cell type in the interface were investigated here. Immunostaining demonstrated CD4 expression in 80% of the 15 cases tested while CD8 expression was present in 60% of the cases. Polymerase chain reaction (PCR) detected IFN- mRNA expression in 75% of eight cases tested; in contrast IL-10 mRNA was only demonstrated in 50% of these same cases. Proteins extracted from another eight cases of revision tissue were analyzed using Western blotting for IL-17, fractalkine (Fkn) and CD40. IL-17 and Fkn were a consistent feature of all cases tested (8/8), while CD40 was undetectable in one case (7/8). These results show that T cells present in the interface are more commonly of the helper T cell phenotype, although cytotoxic T cells are also present. Helper T cells (Th) are responsible for the polarization of the immune response through their production of key mediators. The PCR results obtained in this study suggest that a Th1 response characterized by the production of IFN- predominates over the Th2, IL-10 mediated response. Furthermore the demonstration of the expression of IL-17, Fkn and CD40, all of which are Th1 associated molecules, supports this conclusion.     

Efficacy of green nanoparticles against cancerous and normal cell lines: a systematic review and meta‐analysis

This study aimed to perform a systematic review and meta‐analysis of papers discussing the efficacy of microbial synthesised metallic nanoparticles (MNPs) against cancerous and normal cell lines by exploiting Bayesian generalised linear (BGL) model. Data was systematically collected from published papers via Cochrane library, Web of Science, PubMed, Science Direct, ProQuest, Scopus, and Embase. Impressively, most of the studies were carried out on HeLa and A549 cancer cell lines. Specifically, a hefty 65.67% of studies employed bacteria to biofabricate MNPs. Significantly, BGL meta‐analysis represented highly valuable information. Hence, based on adjusted analysis, the MNPs with the size of 25–50 nm were found to be far less cytotoxic than the MNPs with the size of ≤25 nm (OR = 0.233, P  ˂ 0.05) against either cancerous or normal cell lines. Interestingly, it was found that the odds of cytotoxicity in cancerous cell lines were practically nine times more than normal cell lines, representing the substantially more cytotoxicity of MNPs in cancerous cell lines (OR = 9.004, P  ˂ 0.001). Green MNPs mentioned here may be developed as novel anti‐cancer agents, which could lead to a revolution in the treatment of cancer.Inspec keywords: reviews, nanoparticles, cancer, nanomedicine, magnetic particles, nanomagnetics, nanofabrication, cellular biophysics, Bayes methods, microorganisms, toxicology, antibacterial activityOther keywords: green nanoparticles, cancerous cell lines, normal cell lines, review, meta‐analysis, microbial synthesised metallic nanoparticles, Bayesian generalised linear model, HeLa cancer cell lines, A549 cancer cell lines, gram‐negative bacteria, gram‐positive bacteria, cytotoxicity, anticancer agents, cancer disease treatment     

Multiplexed Biosensing Diagnostic Platforms Detecting Autoantibodies to Tumor-Associated Antigens from Exosomes Released by CRC Cells and Tissue Samples Showed High Diagnostic Ability for Colorectal Cancer

Colorectal cancer (CRC) is the second leading cause of cancer-related death worldwide. The five-year survival rate of CRC patients depends on the stage at diagnosis, being higher than 80% when CRC is diagnosed in the early stages but lower than 10% when CRC is diagnosed in advanced stages. Autoantibodies against specific CRC autoantigens (tumor-associated antigens (TAAs)) in the sera of patients have been widely demonstrated to aid in early diagnosis. Thus, we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy. To that end, we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis. A total of 103 proteins were identified as potential autoantigens specific to CRC. After bioinformatics and meta-analysis, we selected 15 proteins that are more likely to be actual CRC autoantigens in order to evaluate their role in CRC prognosis by Western blot (WB) and immunohistochemistry (IHC). We found dysregulation at the protein level for 11 of these proteins in both tissue and plasma exosome samples from patients, along with an association of nine of these proteins with CRC prognosis. After validation, all but one showed a statistically significant high diagnostic ability to distinguish CRC patients and individuals with premalignant lesions from healthy individuals, either by luminescence Halotag-based beads, or by a multiplexed biosensing platform involving the use of magnetic microcarriers as solid support modified with covalently immobilized Halotag fusion proteins constructed for CRC detection. Taken together, our results highlight the usefulness of the approach defined here to identify the TAAs specific to chronic diseases; they also demonstrate that the measurement of autoantibody levels in plasma against the TAAs identified here could be integrated into a point-of-care (POC) device for CRC detection with high diagnostic ability.     

Targeted Theranostic Nano Vehicle Endorsed with Self‐Destruction and Immunostimulatory Features to Circumvent Drug Resistance and Wipe‐Out Tumor Reinitiating Cancer Stem Cells

The downsides of conventional cancer monotherapies are profound and enormously consequential, as drug‐resistant cancer cells and cancer stem cells (CSC) are typically not eliminated. Here, a targeted theranostic nano vehicle (TTNV) is designed using manganese‐doped mesoporous silica nanoparticle with an ideal surface area and pore volume for co‐loading an optimized ratio of antineoplastic doxorubicin and a drug efflux inhibitor tariquidar. This strategically framed TTNV is chemically conjugated with folic acid and hyaluronic acid as a dual‐targeting entity to promote folate receptor (FR) mediated cancer cells and CD44 mediated CSC uptake, respectively. Interestingly, surface‐enhanced Raman spectroscopy is exploited to evaluate the molecular changes associated with therapeutic progression. Tumor microenvironment selective biodegradation and immunostimulatory potential of the MSN‐Mn core are safeguarded with a chitosan coating which modulates the premature cargo release and accords biocompatibility. The superior antitumor response in FR‐positive syngeneic and CSC‐rich human xenograft murine models is associated with a tumor‐targeted biodistribution, favorable pharmacokinetics, and an appealing bioelimination pattern of the TTNV with no palpable signs of toxicity. This dual drug‐loaded nano vehicle offers a feasible approach for efficient cancer therapy by on demand cargo release in order to execute complete wipe‐out of tumor reinitiating cancer stem cells.     

Digital detection of multiple minority mutants in stool DNA for noninvasive colorectal cancer diagnosis

Somatic mutations in stool DNA are quite specific to colorectal cancer (CRC), but a method being able to detect the extraordinarily low amounts of mutants is challengeable in sensitivity. We proposed a hydrogel bead-array to digitally count CRC-specific mutants in stool at a low cost. At first, multiplex amplification of targets containing multiple mutation loci of interest is carried out by a target enriched multiplex PCR (Tem-PCR), yielding the templates qualified for emulsion PCR (emPCR). Then, after immobilizing the beads from emPCR on a glass surface, the incorporation of Cy3-dUTP into the mutant-specific probes, which are specifically hybridized with the amplified beads from emPCR, is used to color the beads coated with mutants. As all amplified beads are hybridized with the Cy5-labeled universal probe, a mutation rate is readily obtained by digitally counting the beads with different colors (yellow and red). A high specificity of the method is achieved by removing the mismatched probes in a bead-array with electrophoresis. The approach has been used to simultaneously detect 8 mutation loci within the APC, TP53, and KRAS genes in stools from eight CRC patients, and 50% of CRC patients were positively diagnosed; therefore, our method can be a potential tool for the noninvasive diagnosis of CRC.     

Identification of low-abundance cancer biomarker candidate TIMP1 from serum with lectin fractionation and peptide affinity enrichment by ultrahigh-resolution mass spectrometry

As investigating a proteolytic target peptide originating from the tissue inhibitor of metalloproteinase 1 (TIMP1) known to be aberrantly glycosylated in patients with colorectal cancer (CRC), we first confirmed that TIMP1 is to be a CRC biomarker candidate in human serum. For this, we utilized matrix-assisted laser desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) showing ultrahigh-resolution and high mass accuracy. This investigation used phytohemagglutinin-L(4) (L-PHA) lectin, which shows binding affinity to the β-1,6-N-acetylglucosamine moiety of N-linked glycan on a protein, to compare fractionated aberrant protein glycoforms from both noncancerous control and CRC serum. Each lectin-captured fraction containing aberrant glycoforms of TIMP1 was digested by trypsin, resulting in the tryptic target peptide, representative of the serum glycoprotein TIMP1. The resulting target peptide was enriched using a stable isotope standard and capture by the antipeptide antibody (SISCAPA) technique and analyzed by a 15 T MALDI FTICR mass spectrometer with high mass accuracy (Δ < 0.5 ppm to the theoretical mass value of the target peptide). Since exact measurement of multiplex isotopic peaks of the target peptide could be accomplished by virtue of high mass resolution (Rs > 400,000), robust identification of the target peptide is only achievable with 15 T FTICR MS. Also, MALDI data obtained in this study showed that the L-PHA-captured glycoforms of TIMP1 were measured in the pooled CRC serum with about 5 times higher abundance than that in the noncancerous serum, and were further proved by MRM mass analysis. These results confirm that TIMP1 in human serum is a potent CRC biomarker candidate, demonstrating that ultrahigh-resolution MS can be a powerful tool toward identifying and verifying potential protein biomarker candidates.     

Magnetic cell separation using antibody binding with protein a expressed on bacterial magnetic particles

Bacterial magnetic particles (BacMPs) are efficient platforms of proteins for surface display systems. In this study, mononuclear cells from peripheral blood were separated using BacMPs expressing protein A on the BacMP membrane surface (protein A-BacMPs), which were complexed with the Fc fragment of anti-mouse IgG antibody. The procedure of positive selection involves incubation of mononuclear cells and mouse monoclonal antibodies against different cell surface antigens (CD8, CD14, CD19, CD20) prior to treatment with protein A-BacMP binding with rabbit anti-mouse IgG secondary antibodies. Flow cytometric analysis showed that approximately 97.5 +/- 1.7% of CD19(+) and CD20(+) cells were involved in the positive fraction after magnetic separation. The ratio of the negative cells in the negative fraction was approximately 97.6 +/-1.4%. This indicates that CD19(+) and CD20(+) cells can be efficiently separated from mononuclear cells. Stem cell marker (CD34) positive cells were also separated using protein A-BacMP binding with antibody. May-Grunwald Giemsa stain showed a high nuclear/cytoplasm ratio, which indicates a typical staining pattern of stem cells. The separated cells had the capability of colony formation as hematopoietic stem cells. Furthermore, the inhibitory effect of magnetic cell separation on CD14(+) cells was evaluated by measurement of cytokine in the culture supernatant by ELISA when the cells were cultured with or without lipopolysaccharide (LPS). The induction of IL1-beta, TNFalpha, and IL6 was observed in the presence of 1 ng/mL LPS in all fractions. On the other hand, in the absence of LPS, BacMPs had little immunopotentiation to CD14(+) cells as well as that of artificial magnetic particles, although TNFalpha and IL6 were slightly induced in the absence of LPS in the positive fraction.     

Exosomal CD44 Cooperates with Integrin α6β4 to Support Organotropic Metastasis via Regulating Tumor Cell Motility and Target Host Cell Activation

Rapid metastasis to vital organs such as the lung, liver, and brain is responsible for the vast majority of pancreatic cancer deaths. Liver metastasis of pancreatic cancer accounts for the high mortality rate in patients. Exosomes derived from pancreatic cancer cells tend to be enriched in proteins that are anchored to the cell membrane, supporting the reprogramming of the tumor microenvironment and the progression of distant metastatic lesions. For the first time, our study has demonstrated that cluster of differentiation 44 (CD44), a transmembrane glycoprotein delivered by exosomes, is involved in the metastatic process of pancreatic cancer. Moreover, CD44 was found to interact with integrin α6β4 to form a complex, thereby remodeling intracellular skeleton proteins, and to promote tumor cell motility through the activation of the Src and Ras signaling cascades. Notably, we also demonstrated that the CD44–α6β4 complex can be delivered to the target region via the paracrine effects of exosomes. The selective uptake of CD44-competent tumor exosomes by liver cells activated fibrotic pathways and generated a pre-metastatic niche by stimulating the cytokines, proinflammatory factors, and growth factors that ultimately support tumor metastasis. Our results suggest the potential application of exosomal CD44 as a biomarker for the clinical diagnosis of and therapy for pancreatic cancer.     

Characterization of surface-modified nanoparticles for in vivo biointeraction. A sedimentation field flow fractionation study

Sedimentation field flow fractionation (SdFFF) is an emerging high-performance analytical tool for separation and determination of size and adsorption characteristics of colloidal particles. This study demonstrates how SdFFF can be used to characterize nanoparticles prepared for in vivo applications including (1) the quantification of polymer uptake on nanoparticles where surface coverage is crucial and (2) the coupling of cell adhesive peptides containing the Arg-Gly-Asp motif (RGD). Quantitative information about polymer adhesion in order to prepare a bioinert surface and an accurate determination of ligand uptake are both of obvious importance for the understanding of, for example, relations between the number of attached molecules for biointeraction and an observed therapeutic effect. In addition, the present work highlights the necessity to perform careful characterization of commercially available particulate starting materials, in terms of size and polydispersity, prior to biological experimentation.     

Open-tubular capillary cell affinity chromatography: single and tandem blood cell separation

In this paper, an open-tubular capillary cell affinity chromatography (OT-CAC) method to enrich and separate target cells is described. Open tubular capillaries coated with anti-CD4, anti-CD14, or anti-CD19 antibodies were used as affinity chromatography columns to separate target blood cells. Cells were eluted using either shear force or bubbles. Bubbles were used to elute the captured cells without diluting the captured cells appreciably, while maintaining viability (the viability of the recovered cells was 85.83 +/- 7.34%; the viability of the cells was 90.41 +/- 3.49% before separation). Several aspects of the OT-CAC method were studied, such as the affinity of one antibody between two different cell lines, the effect of shear force, and the recovery of captured cells. Single- and multicell type separations were demonstrated by isolating CD4+ cells with antiCD4 coated capillary and isolating CD4+ and CD19+ cells with two capillaries in tandem from blood samples. In the one cell type isolation test, an average of 87.7% of the recovered cells from antiCD4 capillary were lymphocytes and an average of 97.7% of those lymphocytes were CD4+ cells. In the original blood sample, only 14.2% of the leukocytes were CD4+ cells. Two capillary columns were also run in tandem, separating two blood cell types from a single sample with high purity. The use of different elution shear forces was demonstrated to selectively elute one cell type. This method is an inexpensive, rapid, and effective method to separate target cells from blood samples.