Research On The Detection Method Of Cell Proliferation Ability Based On Delayed Luminescence Kinetics

Human Umbilical Cord Mesenchymal Stem Cells(hUC-MSCs)have a high degree of differentiation and self-replication ability,and have broad application prospects in the fields of bioremediation,diabetes,and immunological disease treatment.Whether it is used for cell culture or cell-based therapy,the proliferation ability of stem cells is an important factor that needs to be considered and evaluated.Although there are many biochemical methods that can detect the proliferation ability of cells,most of these methods are invasive and cause irreversible damage to cells,making it difficult to meet the requirements of stem cell production and clinical applications.Photo-induced delayed luminescence(DL)contains rich life information,and has sensitive indicators for changes in cell physiological and pathological conditions,and is expected to become a new method for rapid and non-invasive detection of cell proliferation.This paper uses DL technology to study the proliferation ability of hUC-MSCs,aiming to find out the parameters that can quantitatively characterize the proliferation ability.The research work of the thesis mainly includes four parts:Firstly,the basic characteristics of DL from hUC-MSCs were studied,including five aspects:cell sedimentation,light intensity,light time,light wavelength,and the influence of cell number.Based on the coherence theory,a dynamic model of DL decay was derived and established.The results show that compared with the traditional hyperbolic decay model,the model established in this paper fits the DL decay curve of high and low viability cells better,which proves that the model can be applied to the quantitative characterization of the DL characteristics of hUC-MSCs.Secondly,the DL decay kinetic model was used to detect cell viability.Compared with high viability cells,the parameterN_s/N₀ of low viability cells decreased by 43%.After T test analysis,the P value of the high and low vitality parameterN_s/N₀ is 1.2E-4,and the difference between the two is statistically significant.With the decrease of cell viability,the parameterN_s/N₀ showed a monotonous decrease.After K-means cluster analysis,the reference threshold for distinguishing cell viability is 24.32.Thirdly,the DL decay kinetics model is used to detect the proliferation ability of highly viable cells.Select cell lines with different proliferation ability to analyze the DL decay kinetics.The results show that the parameterN₀×(?-G)is closely related to the proliferation ability.Compared with cells with strong proliferation ability,the net number of photonsN₀×(?-G)dissipated in the photon field of cells with weak proliferation ability at the initial moment increased by 1.5 times.After T test analysis,the P value is 1.5E-3,and the difference between the two is statistically significant.Through K-means clustering analysis,it is proposed that the reference threshold for distinguishing strong and weak proliferation ability is 3.18 count/ms,and the parameterN₀×(?-G)is expected to be an indicator of quantitative characterization and evaluation of proliferation ability.Finally,realize the algorithm design and program development for detecting hUC-MSCs proliferation ability.The algorithm mainly includes five parts:data loading,automatic parameter fitting,high and low viability analysis,strong and weak proliferation ability analysis and result display.After testing,the algorithm runs stably,which provides a new idea for rapid and non-destructive detection of cell proliferation.