Systems and methods to determine the age of cells

一种细胞、红血细胞的技术，应用在颗粒和沉降分析、仪器、分析材料等方向，能够解决未显示出红细胞寿命或平均年龄的确定等问题

Active Publication Date: 2017-05-24
BECKMAN COULTER INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of these techniques have been shown to consistently provide a determination of the lifespan or mean age of red blood cells

Method used

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  • Systems and methods to determine the age of cells
  • Systems and methods to determine the age of cells
  • Systems and methods to determine the age of cells

Examples

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example 1

[0205] An assay is described that measures the cellular hemoglobin and HbA1c percentages of blood samples and fractionates the cells into reticulocytes and ten fractions of mature erythrocytes. Glucose and hemoglobin react irreversibly together to form HbA1c (Amadori product). This response is a continuous phenomenon as a function of time and glucose concentration. The formation of HbA1c is a controlling element of blood sugar balance in diabetic patients. Normal value of HbA1c: 5% of total hemoglobin.

[0206] Materials and Methods .

[0207] The following materials were used in this assay:

[0208] 1. Blood samples treated with 0.7 mM ethylenediaminetetraacetic acid (EDTA) as an anticoagulant and stored at 4°C for no more than ten days.

[0209] 2. Spheroidizing reagent: 0.137M sodium chloride, 0.005M HEPES, 0.0005M (D+) trehalose, 0.083M formaldehyde, 0.04mM n-dodecyl β-D-maltoside, 0.5ml / l Proclin-300, 0.5 mg / l acridine orange and a certain amount of sodium hydroxid...

example 2

[0224] Percentage HbA1c data from a reference laboratory using three different assays compared to a flow cytometry procedure Comparison of the correlation curves of HbA1c percentage data derived from the sequence .

[0225] Percent HbA1c was determined on 120 samples by reference laboratories using three different HbA1c assay methods; (Primus Ultra2: affinity chromatography, Roche Unimate: immunoturbidimetry, and Tosoh G7 variant: cation exchange chromatography). Flow cytometry percent HbAlc values ​​were determined in the presence of reference control cells of known percent HbAlc. This document describes the conversion of arbitrary HbA1c percent units to percent HbA1c values.

[0226] Methods and Results

[0227] In the first case, we show that fluorescently labeled antibodies have free access to HbA1c molecules inside immobilized erythrocytes. Red blood cells are fixed with formaldehyde and lysed in water and further disintegrated in the presence of detergent. The l...

example 3

[0244] Data from six normal subjects; representation of FL4 / SS values ​​and corrected FL4 / SS values ​​in different fractions, and conversion of percentage HbA1c to time in each fraction .

[0245] This example presents data from six normal subjects and the conversion of fraction limit FL4 / SS values ​​to time. For reasons of comparison with blood samples taken from diabetic individuals, in the first method, blood samples were selected that were representative of normal non-diabetic individuals who were considered to experience a constant blood glucose concentration. To further confirm that the parameters derived from normal samples represent constant blood glucose concentrations, the parameters of 6 normal samples were averaged.

[0246] Raw data generated by flow cytometry were used to fractionate mature cell populations into fractions containing equal numbers of cells as described in Example 1 . Cells were organized on the HbA1c percentage axis according to their increas...

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Abstract

Systems and methods are provided for determining the age of cellular hemoglobin in individual red blood cells in a blood sample by determining the percentage of HbA1c. In embodiments, a method includes measuring side scatter and fluorescence of the individual red blood cells and identifying immature red blood cells and mature red blood cells from the side scatter and fluorescence measurement. In embodiments, data collected includes the exact number of red blood cells, the fraction limits in fluorescence and side scatter units, the mean value of each fraction in fluorescence and side scatter units, the mean FL1 values per fraction in arbitrary units and the mean side scatter values per fraction in arbitrary units. In embodiments, a method also includes deriving an HbA1c content from the measured mean fluorescence of the individual red blood cells and determining the percentage of HbA1c from the HbA1c content and the hemoglobin content of the red blood cells.

Description

[0001] Cross references to related patent applications [0002] This application was filed as a PCT International Patent Application on September 11, 2015, and claims priority to U.S. Patent Application Serial No. 62 / 059,684, filed on October 3, 2014, and claims Priority to US Patent Application Serial No. 62 / 049,970, the disclosures of both patent applications are incorporated herein by reference in their entirety. technical field [0003] The present disclosure relates to systems and methods for determining the age of cellular hemoglobin and / or the average age of red blood cells in a cell. Background technique [0004] Red blood cell longevity has been derived by measuring the in vivo clearance of chromium-51-labeled red blood cells (see Gray, S.J. and Stirling, K. 1950, J. Clin. Invest. 29:1604-1613 (Gray, S.J. , 1950, Review of Journal of Clinical Investigation, Vol. 29, pp. 1604-1613); Dacie and Lewis; Practical Haematology, Ninth Edition, 2001, Ed.; S.M. Lewis, B.J. B...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72G01N15/14
CPCG01N33/721G01N33/726G01N2333/805G01N2015/1488G01N2015/1402G01N15/1459G01N2015/1477G01N33/723G01N33/492G01N2015/014G01N33/582G01N33/80
Inventor 安德里斯·范艾格斯霍芬法布里斯·马莱格克里斯托夫·戈德弗罗伊恩里克·拉贝力诺
Owner BECKMAN COULTER INC
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