Background. Mesenchymal stem cells (MSCs) are currently exploited in numerous clinical trials to investigate their potential in immune regulation, hematopoesis or tissue regeneration. The most common source of MSCs for clinical use is human bone marrow. To generate sufficient numbers of cells relevant to clinical use in most cases the high volumes (20-50 ml) of bone marrow aspirates are taken. Methods. In this pilot study, 8 healthy bone marrow donors were included. Two different MSC extraction methods were evaluated: MSCs extraction from 60 ml of bone marrow using density gradient and MSCs extraction from 6 ml using red blood cell (RBC) lysis. Results. Our results showed that after RBC lysis the efficient amount of human MSCs can be isolated from 10 times less bone marrow volume (6 ml). Moreover, using small volume of bone marrow the adequate therapeutical dose of MSCs could be achieved during similar period of time (3-4 weeks). In conclusion, we have shown that MSCs isolation using RBC lysis is an effective and more advantageous method in comparison to standard MSCs isolation using density-gradient. Using RBC lysis from small volume of bone marrow the same amount of MSCs were obtained as usually using large volume and density-gradient.
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Cervical cancer remains an important cause of women morbidity and mortality. The progression of cervical pathology correlates with the HPV integration into the host genome. However, the data on the viral integration status in cervical dysplasias are controversial. The aim of the current study was to evaluate the status of HPV integration in two types of cervical pathology – invasive and non invasive cervical cancer (e.g. carcinoma in situ). 156 women were included in the study: 66 women were diagnosed with invasive cervical cancer (CC) and 90 with non invasive cervical cancer (carcinoma in situ, CIS). 74.2% [95% PI: 63.64÷84.76] of specimens collected from women with diagnosed CC and 85.6% [95% PI: 85.53÷92.85] of CIS specimens were positive for HPV. The most prevalent HPV genotype in both groups was HPV16. To evaluate HPV integration, three selected HPV16 E2 gene fragments were analyzed by PCR. In the majority of CC and CIS specimens the amplification of all three HPV16 E2 gene fragments was observed. The episomal HPV16 form was detected in the majority of CC and CIS specimens. The deletion of all three HPV16 E2 gene fragments was detected in 9.4% of CC specimens and 2.2% of CIS specimens. Finally, integration status could not be used as diagnostical additional test to distinguish between invasive and non invasive cervical cancer.
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In some countries the cervical cancer incidence and mortality rates are much higher compared to the European average. The differences of HPV and its type prevalence between countries and regions influence cervical cancer incidence and mortality. Regarding the differences in cervical cancer incidence and mortality in Lithuania and Belarus, the aim of this study was to describe HPV infection level and HPVs type distribution among two study groups of patients with moderate or severe cervical intraepithelial neoplasia (CIN2-3) and cervical cancer. Our data shows that 74.2% [95% CI: 63.64÷84.76] of Lithuanian patients with cervical cancer and 85.6% [95% CI: 85.53÷92.85] of the study group with CIN2-3 were HPV positive, while in the study groups of Belarusian patients HPV infection was detected in 92.6% [95% CI, 74.25÷98.71] and 65.4% [95% CI, 44.36÷82.06] cases respectively. HPV 16 was the most prevalent type in Lithuanian as well as in Belarusian patients of the study groups. HPV 18 in Lithuanian patients of the study group with cervical cancer was identified in 10.2% [95% CI: 1.73÷18.67] and in the study group with CIN2-3 - in 2.6% [95% CI: 0.95÷6.15] of cases. HPV 18 was not detected in Belarusian patients of both groups.
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