Culture determination of human embryonic lung diploid cells and analysis on their sensitivity to encephalomyocarditis virus
10.13200/j.cnki.cjb.004685
- VernacularTitle:人胚肺二倍体细胞的培养检定及其对脑心肌炎病毒敏感性的分析
- Author:
Chen LIANG
- Publication Type:Journal Article
- Keywords:
Human embryonic lung diploid cells;
Encephalomyocarditis virus(EMCV);
Biological characteristics;
Safety;
Viral sensitivity
- From:
Chinese Journal of Biologicals
2026;39(04):403-415
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify and verify five strains of human embryonic lung diploid cells independently isolated and cultured, and to explore their sensitivity to encephalomyocarditis virus(EMCV), so as to screen for promising human embryonic lung diploid cells with application prospects.MethodsFive human embryonic lung cell strains were successfully isolated from artificially aborted embryos using the trypsin digestion method and named MU-L6, MU-L9, MU-L10, MU-L11 and MUL12. The five cell strains were identified by growth curve analysis, chromosomal karyotype analysis, immunofluorescence,nucleic acid electrophoresis, and short tandem repeat(STR) profiling. Their safety was assessed through sterility testing,mycoplasma detection, examination for endogenous and exogenous viral agents, and tumorigenicity testing. The sensitivity of the five cell strains to EMCV was determined by the 50% tissue culture infective dose(TCID_(50)) assay.ResultsAfter continuous subculturing, the MU-L10 cell strain could be passaged up to 81 generations, while the other four strains reached between 30 and 45 generations. The maximum proliferation density of MU-L10 cells was(44. 07 ± 2. 57) × 10~4 cells/mL, with a population doubling time of(25. 77 ± 1. 00) hours. All five human embryonic lung diploid cell strains exhibited fibroblast-like morphology and diploid karyotypes. No interspecies cross-contamination was detected. MU-L9, MU-L10, MU-L11 and MUL12 possessed unique cellular identities, indicating they are novel cell lines. Safety evaluations revealed no contamination with exogenous bacteria or viruses in any of the five cell strains. MU-L6, MU-L9, MU-L10 and MU-L11 were free of mycoplasma contamination. No colony formation was observed in the in vitro tumorigenicity assay. All five cell strains demonstrated sensitivity to EMCV infection. At 48 hours post-infection, the TCID_(50)of MU-L10 cells was 10~((-8.22 ± 0.14))/0.1 mL,showing no significant difference compared to the control MRC-5 cells(P = 0. 317 7).ConclusionThe independently isolated and cultured MU-L10 cell strain exhibits high passage longevity, excellent biosafety and genetic stability, making it a promising candidate for research on viral vaccines.
