|
|
Cell Growth and Drug Effect Differences Induced by Inoculum Number Variations and Comparison of Cell Counting Methods |
XUE Zhi-chao1,ZENG Jia-ming1,LI Yong-shu2,ZHAO Jia-wei1,ZHAO Yang1,DAI Xin-hua1,GONG Xiao-yun1 |
1. Technology Innovation Center of Mass Spectrometry for State Market Regulation, Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China
2. Shenzhen Institute for Technology Innovation, National Institute of Metrology, Shenzhen,Guangdong 518055, China |
|
|
Abstract Hela cells were used to investigate the changes in cell growth rate and the influence of drug effect induced by the cell number differences at the cell seeding step. The experimental results indicated that the larger the initial number of cells, the higher the density after adhesion, the faster the growth rate, and the lower the inhibitory effect of drugs on its growth. Therefore, the accuracy and efficiency of cell counting are of great importance for in vitro studies. Three commonly used cell counting methods were compared including the cell counting plate method, the cell counting apparatus method, and the flow cytometry method. Their linear regression R2 were 0.9835, 0.9997, and 0.9960 respectively. Under the same condition, the cell counting apparatus method held the highest accuracy. In addition, indirect methods such as crystal violet staining, basal area photography, protein content estimation, and protein reference GAPDH measurement were also evaluated in to provide data reference for cell number measuring in biological experiments.
|
Received: 04 July 2022
Published: 08 March 2023
|
|
|
|
|
[16] |
Balsam J, Bruck H A, Rasooly A. Mobile flow cytometer for mHealth[J]. Methods Mol Biol, 2015, 1256: 139-153.
|
[21] |
Vembadi A, Menachery A, Qasaimeh M A. Cell Cytometry: Review and Perspective on Biotechnological Advances[J]. Front Bioeng Biotechnol, 2019, 7:147.
|
[22] |
刘瑛颖, 张玲, 杨乔斯. 尿沉渣白细胞计数标准物质的研制[J]. 计量学报, 2022. 43(3): 427-432.
|
|
Liu J M, Jia X Q, Di B. Application of three-dimensional cell model in tumor research[J]. J China Pharm Univ, 2020, 51(2): 152-160.
|
[4] |
Faruqui N, Kummrow A, Fu B, et al. Cellular Metrology: Scoping for a Value Proposition in Extra-and Intracellular Measurements[J]. Front Bioeng Biotechnol, 2020,7: 456.
|
[6] |
Amoozadeh Y, Anwer S, Dan Q, et al. Cell confluence regulates claudi-2 expression: possible role for ZO-1 and Rac[J]. Am J Physiol Cell Physiol, 2018, 314(3): 366-378.
|
[11] |
李晶晶, 罗明志, 杨莹, 等. 三种细胞计数法的对比试验研究[J]. 江西科技师范大学学报, 2016(6): 79-81.
|
[10] |
Araujo R P, Liotta L A, Petricoin E F. Proteins, drug targets and the mechanisms they control: the simple truth about complex networks[J]. Nature Reviews Drug Discovery, 2007, 6(11): 871-880.
|
[12] |
Mirabelli P, Coppola L, Salvatore M. Cancer Cell Lines Are Useful Model Systems for Medical Research[J]. Cancers (Basel), 2019,11(8):1098.
|
[14] |
Xu G, Pei Q Y, Ju C G, et al. Detection on effect of different processed Cibotium barometz on osteoblasts by CCK-8[J]. China Journal of Chinese Materia Medicai, 2013. 38(24): 4319-4323.
|
[18] |
Kobayashi H, Takemura Y, Ohnuma T. Relationship between Tumor-Cell Density and Drug Concentration and the Cytotoxic Effects of Doxorubicin or Vincristine-Mechanism of Inoculum Effects[J]. Cancer Chemotherapy and Pharmacology, 1992, 31(1): 6-10.
|
|
Liu Y Y, Zhang L, Yang Q S. Development of reference material for white blood cell count in urine sediment.[J]. Acta Metrologica Sinica, 2022, 43(3): 427-432.
|
[25] |
Trajkovic K, Valdez C, Ysselstein D, et al. Fluctuations in cell density alter protein markers of multiple cellular compartments, confounding experimental outcomes[J]. PLoS One, 2019, 14(2): 0211727.
|
[5] |
Abe M, Havre P A, Urasaki Y, et al. Mechanisms of confluence-dependent expression of CD26 in colon cancer cell lines[J]. BMC Cancer, 2011, 11: 51.
|
[13] |
Bitar M, Brown R A, Salih V, et al. Effect of Cell Density on Osteoblastic Differentiation and Matrix Degradation of Biomimetic Dense Collagen Scaffolds[J]. Biomacromolecules, 2008, 9(1): 129-135.
|
[3] |
Amelian A, Wasilewska K, Megias D, et al. Application of standard cell cultures and 3D in vitro tissue models as an effective tool in drug design and development[J]. Pharmacological Reports, 2017, 69(5): 861-870.
|
|
Li J J, Luo M Z, Yang Y, et al. Comparison of Three Cell Count Methods[J]. Journal of Jiangxi Science & Technology Normal University, 2016(6): 79-81.
|
[20] |
Bena E C, Giudice D M. Initial cell density encodes proliferative potential in cancer cell populations[J]. Sci Rep, 2021, 11(1): 6101-6111.
|
[2] |
Wong C H, Siah K W, Lo A W. Estimation of clinical trial success rates and related parameters[J]. Biostatistics, 2019, 20(2): 273-286.
|
[7] |
Poumay Y, Pittelkow M R. Cell density and culture factors regulate keratinocyte commitment to differentiation and expression of suprabasal K1/K10 keratins[J]. J Invest Dermatol, 1995, 104(2): 271-276.
|
[8] |
Nusinow D P, Szpyt J, Ghandi M, et al. Quantitative Proteomics of the Cancer Cell Line Encyclopedia[J]. Cell, 2020, 180(2): 387-402.
|
[15] |
Chiba K, Kawakami K, Tohyama K. Simultaneous evaluation of cell viability by neutral red, MTT and crystal violet staining assays of the same cells[J]. Toxicol In Vitro, 1998, 12(3): 251-258.
|
[19] |
Greene J M, Levy D, Herrada S P, et al. Mathematical Modeling Reveals That Changes to Local Cell Density Dynamically Modulate Baseline Variations in Cell Growth and Drug Response[J]. Cancer Res, 2016, 76(10): 2882-2890.
|
[23] |
王斌, 隋志伟, 刘思渊, 等. 基于流式分析技术的奶粉中金黄色葡萄球菌活菌快速定量检测方法研究[J]. 计量学报,2021,42(2): 250-258.
|
|
Wang B, Sui Z W, Liu S Y, et al. Rapid and quantitative detection of viable staphylococcus aureus.in milk powder based on flow cytometry technology[J]. Acta Metrologica Sinica, 2021, 42(2): 250-258.
|
[1] |
刘佳敏, 贾晓青, 狄斌.三维细胞模型在肿瘤研究中的应用[J]. 中国药科大学学报,2020,51(2):152-160.
|
[9] |
Garnett M J, Edelman E J, Heidorn S J, et al. Systematic identification of genomic markers of drug sensitivity in cancer cells[J]. Nature, 2012, 483(7391): 570-575.
|
[17] |
王真真. 1,25(OH)2D3联合顺铂对宫颈癌Hela细胞增殖及P27蛋白表达的影响[D]. 承德:承德医学院,2017.
|
[24] |
Wu Y, Wu M, He G, et al. Glyceraldehyde-3-phosphate dehydrogenase: A universal internal control for Western blots in prokaryotic and eukaryotic cells[J]. Analytical Biochemistry, 2012, 423(1): 15-22.
|
[26] |
Liang F, Wen X, Geng Y, et al. Growth Rate and Biomass Productivity of Chlorella as Affected by Culture Depth and Cell Density in an Open Circular Photobioreactor[J]. Journal of Microbiology and Biotechnology, 2013, 23(4): 539-544.
|
|
|
|