Contributor:

Sypecka, Joanna (Promotor) ; Ziemka-Nałęcz, Małgorzata (Promotor pomocniczy)

Publisher:

Instytut Medycyny Doswiadczalnej i Klinicznej im. Miroslawa Mossakowskiego PAN

Place of publishing:

Warszawa

Description:

Bibliografia zawiera 225 pozycje ; 113s.: il., wykr., tabl., fotogr.; 30 cm.

Degree name:

doctor

Level of degree:

2

Degree discipline :

medical sciences

Degree grantor:

Instytut Medycyny Doświadczalnej i Klinicznej im. M. Mossakowskiego PAN

Abstract:

Perinatal asphyxia (neonatal hypoxia-ischemia, HI) is associated with abnormal blood flow and impaired gas exchange during the perinatal period. The high mortality rate or motor and cognitive disabilities make complications after HI a significant social problem. In developed countries, the problem affects 1-8 per 1,000 live births. Currently, the only available therapy is therapeutic hypothermia. Despite multidirectional research, an effective neuroprotective and neuroregenerative strategy is still lacking.The interrelationship between glial cells in the developing brain following the temporary glucose and oxygen deprivation is poorly understood. Among glial cells, there are macroglia, including astrocytes and oligodendrocytes (OL), and microglia. Glial cells are involved in the normal development and functioning of the nervous system, in maintaining homeostasis in the tissue microenvironment and in the immune response. Activated glial cells produce immunomodulatory factors, including cytokines (pro-inflammatory or anti-inflammatory). The interplay between the cells may be crucial for the predominance of neural repair processes over inflammatory and neurodegenerative processes.The purpose of this study is to analyze the effects of transient oxygen-glucose deprivation (in vitro HI model) on the survival, proliferation, phenotype and morphology of neonatal rat glial cells, and to analyze their secretory profile. In order to mimic conditions in neural tissue, physiological normoxia (5% oxygen concentration) was applied, and selected biomimetic compounds (laminin, fibronectin, ECM Gel) were tested to coat the surfaces of culture dishes. To assess the role of selected factors (trophic factors, cytokines and their receptors) involved in interactions between glial cells, restrictive culture media were used.The study was carried out on glial cells derived from the brains of Wistar rat pups (24-48h postpartum). A primary mixed glial culture was established from the isolated brain hemispheres. After 11-12 days of in vitro culture, individual glial fractions were isolated by sequential shaking. Studies were conducted on cell populations cultured as monofractions and their co-cultures. At selected time points, culture media were collected and the concentration of selected active factors was quantified (ELISA, Luminex).Analysis of the glial cell secretome showed that astrocytes are the main source of IGF-1. For several of the factors studied, a significant modulating effect of paracrine intercellular interactions on their secretion into the culture medium was demonstrated, including the BDNF factor, the level of which in co-cultures of OL and astrocytes is significantly lower than the level of secretion by the mentioned cell fractions cultured separately. A similar effect of co-cultures was observed in the case of CXCL1 chemokine secretion by OL and microglia, where in all experimental variants there was a significant effect of microglia on the secretion of the mentioned chemokine by OL. In the case of CXCR2 and CXCR4 receptors, however, a significant effect of intercellular interactions was observed in co-cultures of OL and astrocytes.In conclusion, the conducted research is novel in the context of developing an efficient method for obtaining and culturing under physiological normoxia all three main types of neonatal rat glial cells. The use of culture media of the restricted composition, which exclude the presence of factors that can modulate the processes under study, makes it possible to use the culture for in vitro modelling of selected perinatal pathophysiological conditions. In our study, the described culture system was used for in vitro modelling of neonatal HI and studying interactions between glial cells on secretion of selected active factors.

Detailed Resource Type:

PhD Dissertations

Resource Identifier:

oai:rcin.org.pl:239192

Source:

IMDiK PAN, sygn. ZS 426 ; click here to follow the link

Language:

pol

Rights:

Creative Commons Attribution BY 4.0 license

Terms of use:

- ; Copyright-protected material. [CC BY 4.0] May be used within the scope specified in Creative Commons Attribution BY 4.0 license, full text available at:

Digitizing institution:

Mossakowski Medical Research Institute PAS

Original in:

Library of the Mossakowski Medical Research Institute PAS

Access:

Open