Monitoring of Intracellular Enzyme Kinetic Characteristics of Peripheral Mononuclear Cells in Breast Cancer Patients

Afrimzon, Elena; Zurgil, Naomi; Shafran, Yana; Sandbank, Judith; Orda, Ruben; Lalchuk, Shlomo; Deutsch, Mordechai

doi:10.1158/1055-9965.epi-03-0153

Cited by 10 publications

(10 citation statements)

References 41 publications

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“…This indicates that this parameter is not a sensitive marker for post-thawing cell status. The quantitative analysis of the individual cell rate of FDA hydrolysis was done by kinetic measurements of fluorescein accumulation and the calculation of the linear slope of FI increase over time [ 23 , 24 ]. Figure 6(A-D) shows the FDA staining of U937 cells at 3 hours after thawing, and the calculated staining rates (Figure 6E ).…”

Section: Resultsmentioning

confidence: 99%

The individual-cell-based cryo-chip for the cryopreservation, manipulation and observation of spatially identifiable cells. II: Functional activity of cryopreserved cells

et al. 2010

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BackgroundThe cryopreservation and thawing processes are known to induce many deleterious effects in cells and might be detrimental to several cell types. There is an inherent variability in cellular responses among cell types and within individual cells of a given population with regard to their ability to endure the freezing and thawing process. The aim of this study was to evaluate the fate of cryopreserved cells within an optical cryo apparatus, the individual-cell-based cryo-chip (i3C), by monitoring several basic cellular functional activities at the resolution of individual cells.ResultsIn the present study, U937 cells underwent the freezing and thawing cycle in the i3C device. Then a panel of vital tests was performed, including the number of dead cells (PI staining), apoptotic rate (Annexin V staining), mitochondrial membrane potential (TMRM staining), cytoplasm membrane integrity and intracellular metabolism (FDA staining), as well as post-thawing cell proliferation assays. Cells that underwent the freezing - thawing cycle in i3C devices exhibited the same functional activity as control cells. Moreover, the combination of the multi-parametric analysis at a single cell resolution and the optical and biological features of the device enable an accurate determination of the functional status of individual cells and subsequent retrieval and utilization of the most valuable cells.ConclusionsThe means and methodologies described here enable the freezing and thawing of spatially identifiable cells, as well as the efficient detection of viable, specific, highly biologically active cells for future applications.

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Section: Resultsmentioning

confidence: 99%

The individual-cell-based cryo-chip for the cryopreservation, manipulation and observation of spatially identifiable cells. II: Functional activity of cryopreserved cells

et al. 2010

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“…Here we report a new functional assay for the evaluation of lymph node involvement in metastasis that may be performed before the traditional procedure of lymph node histological examination. The FDA staining, occurring only in living cells and applicable in their kinetic studies, is known to indicate a specific cell status such as activation [17,18] or apoptosis [19,20], and to indirectly demonstrate the general nonspecific mechanisms involved in cancer development [16,23].…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, recently, we have shown that, in BC patients, the enzymatic activity of peripheral blood cells interacting with autologous tumor antigens, differ remarkably from the enzymatic reactions in healthy individuals. The enzymatic activity was associated with a prognostic status of BC patients [23].…”

Section: Introductionmentioning

confidence: 96%

Intracellular esterase activity in living cells may distinguish between metastatic and tumor-free lymph nodes

Afrimzon

Deutsch

Shafran

et al. 2008

Clin Exp Metastasis

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“…The specific tumor-associated antigens induce mitogenic stimulation of peripheral lymphocytes resulting in intracellular microviscosity changes, which are manifested in a characteristic kinetic behavior. Moreover, these changes in microviscosity precede other alterations due to activation, such as gene, receptor, and protein expression [ 9 ]. The present study was to articulate the spleenocytes capase3 and granzyme B kinetic behaviors in response to therapeutic vaccination with/without cancer therapeutic adjuvant (creatine analog).…”

Section: Introductionmentioning

confidence: 99%

Implication of caspase-3 and granzyme B expression and activity in spleenocytes of ehrlich ascites carcinoma mice subjected to immunotherapy

Azab¹,

osman²,

El-Fatih³

2011

North Am J Med Sci

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BackgroundCaspase-3 and granzyme B were claimed as apoptotic manipulative enzymes.AimsThe present study was to determine the enzymes expression and activity in cancer and cancer immune therapeutic status and the possible association to cancer common pathological signs targeting the improvement of therapeutic conditions.Material and MethodsMice were immunized with cell lyaste or cell lysate + CKI in the right thigh and challenged with live cells of ehrlich ascites carcinoma (EAC) in the left thigh. The expression and activity of both enzymes in the spleenocytes derived from different subjects (normal, EAC and cell lysate or cell lysate + CKI immunized mice) after cultured with EAC viable cells were determined by colorimetric assay and western blot analysis. In addition, the subjects DNA ladder and serum metalloproteases (MMP 2 and 9) zymography were observed.ResultsThe experimental data revealed over expression of caspase3 and granzyme B in the groups of cell lysate or cell lysate + CKI immunized mice compared to control while down expression were recorded in the EAC subject. The over expression of the 2 enzymes were accompanied with increases in the activities of caspase3 and granzyme B, changes in DNA fragmentation and inhibition of metalloproteases.ConclusionIt could be suggested that, the parameter estimation within the present experimental framework could identify the efficiency of therapeutic vaccine protocols and elucidate the impact of CKI adjuvant with vaccines therapy.

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Monitoring of Intracellular Enzyme Kinetic Characteristics of Peripheral Mononuclear Cells in Breast Cancer Patients

Cited by 10 publications

References 41 publications

The individual-cell-based cryo-chip for the cryopreservation, manipulation and observation of spatially identifiable cells. II: Functional activity of cryopreserved cells

The individual-cell-based cryo-chip for the cryopreservation, manipulation and observation of spatially identifiable cells. II: Functional activity of cryopreserved cells

Intracellular esterase activity in living cells may distinguish between metastatic and tumor-free lymph nodes

Implication of caspase-3 and granzyme B expression and activity in spleenocytes of ehrlich ascites carcinoma mice subjected to immunotherapy

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