Cell preparations to be used in clinical practice must be free of infectious agents. Safety concerns are especially elevated upon the use of human fetal tissues, which are otherwise highly advantageous in cell therapy. We demonstrate that treating fetal samples with antibiotic, extensive washing, and homogenization prior to cryoconservation efficiently removes microbes in general. Screening a large collection by an automatic culture system showed that 89.2% fetal tissue samples were sterile, while contamination was detected in 10.8% samples. Liver and chorion were contaminated more than the brain, kidney, lung, and soft tissues. Broad-range PCR from the bacterial 16s rRNA gene was adopted as a confirmatory assay; however, the concordance between the culture-based and PCR assays was weak. Taxonomic identification was done for contaminated samples by bacteriological methods and sequencing 16s rRNA PCR products. The two approaches revealed different spectra of taxonomic groups sharing only Lactobacillus, the most frequently found genus. In addition, other representatives of vaginal microbiota were detected by culture-based identification, while PCR product sequencing has also revealed a subset of nosocomial microorganisms. Importantly, species known to cause sepsis were identified by both techniques, arguing for their indispensability and mutual complementarity. We suggest that most contaminations are taken up during collection of fetal material rather than originating from an in utero infection. In conclusion, a rigorous microbiological control by culture and PCR is a prerequisite for safe clinical use of fetal tissue suspensions.
To investigate the effect of combined treatment with the inclusion of fetal stem cells (FSCs) on the morphology and functional dynamics of the left ventricle and cognitive functions in patients with chronic heart failure (CHF). Materials and methods: A comparative study was carried out on patients with CHF to examine the effect of combined treatment, including the experimental application of FSCs, on the morphofunctional parameters of the left ventricle and cognitive functions. Patients were examined before FSC treatment (FSCT), and 1 month, 3 months, and 6 months after treatment. The control group consisted of 20 CHF patients of similar age, sex, and New York Heart Association class. Results: It has been proven that FSCs positively affect objective and subjective clinical parameters. A significant reduction of serum type B brain natriuretic peptide was reported as early as 1 month after treatment. Significant increases in the left ventricle ejection fraction and decreases of the end diastolic volume were observed 6 months after treatment. Cognitive performance tests showed improvements on the Mini-Mental State Examination and Frontal Assessment Battery (conceptualization, mental flexibility, programming, sensitivity to interference, inhibitory control, and environmental autonomy) scales. The treatment resulted in significant improvements in the general score and across all cognitive areas of the Mini-Mental State Examination (recall, orientation, attention, calculation, and complex commands) after 3 months, and significant improvements across all Frontal Assessment Battery areas after 6 months. In the control group, these scores showed significant increases only at 6 months after the treatment. In the study group, depression was significantly reduced within 1 month after treatment versus 3 months in the control group. In both groups, reactive anxiety levels dropped after month 3 upon FSCT. FSCT resulted in statistically significant improvements in the contractile activity of the left ventricular myocardium, in cognitive functions, and in the emotional state of CHF patients. Conclusion: The evidence for significant improvements in the contractile function of the left ventricle myocardium, as well as in patients' cognitive and emotional states, was observed in CHF patients after combined treatment with FSC.
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