Mouse Myelinating Oligodendrocyte Differentiation In Vitro
| Autor: | Rebac, Robert, Kužir, Snježana, Alić, Ivan |
|---|---|
| Jazyk: | chorvatština |
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Veterinar Volume 60. Issue 1. |
| ISSN: | 0303-5409 |
| Popis: | Stanice živčanog sustava, neuroni i glija-stanice razvijaju se iz istih matičnih stanica uz izražaj karakterističnih biljega. Glavni cilj ovog istraživanja bio je istražiti diferencijaciju oligodendrocita te analizirati izražaj biljega karakterističnih za pojedini stadij stanica. Istraživanje je provedeno na stanicama izoliranim od novorođenih Thy1 miševa koji pod utjecajem promotora gena Thy1 izražavaju zelenu fluorescentnu bjelančevinu u pojedinim neuronima. Izolirani progenitori oligodendrocita diferencirani su do zrelih, mijelinizirajućih oligodendrocita. Stanice su fiksirane 1., 3., 5. i 7. dan diferencijacije te je na njima učinjena imunocitokemija. Rezultati ovog istraživanja pokazuju da je prvog dana diferencijacije 99 % stanica olig2, sox2 i nestin-pozitivno. Već prvog dana diferencijacije 88 ± 2 % stanica je o4 i o1-pozitivno, dok je svega 10 % stanica gfap-pozitivno. Tijekom diferencijacije stanica udio stanica olig2 smanjuje se do 8 ± 2 %, dok se peti dan diferencijacije pojavljuju zreli mijelinizirajući oligodendrociti koji su pozitivni na mbp i plp. Sedmi dan diferencijacije udio stanica pozitivnih na mbp iznosi 13 ± 2 % stanica, dok udio stanica pozitivnih na plp iznosi 84 ± 3 % stanica. Zaključno, rezultati ovog istraživanja pokazuju da se stanice tijekom sedam dana uzgoja in vitro mogu u potpunosti diferencirati od progenitora oligodendrocita do zrelih, mijelinizirajućih oligodendrocita koji mogu obavljati mijelinizaciju. Cells of the nervous system, neurons, and glia-cells originate from the same stem cell population with the expression of specific markers. The main goal of our study was to analyse the differentiation of oligodendrocytes and show the expression of cell stage-characteristic markers. The study was performed on cells isolated from newborn Thy1 mouse pups. Thy1 mice, under the promoter of the Thy1 gene, express a green fluorescent protein in some neurons. Oligodendrocyte progenitors were differentiated into mature, myelinating oligodendrocytes. The cells were fixed on the 1st, 3rd, 5th and 7th days of differentiation and analysed by immunocytochemistry. Our results showed that on the 1st day of differentiation, 99 % of cells were olig2, sox2, and nestin positive. Moreover, on the 1st day of differentiation 88 ± 2 % of cells were o4 and o1 positive, while only 10 % of cells were gfap positive. During cell differentiation, the proportion of olig2 positive cells decreased to 8 ± 2 %, while on the 5th day of differentiation we observed mature, myelinating oligodendrocytes, mbp, and plp positive cells. On the 7th day of differentiation, 13 ± 2 % of cells were mbp positive, while 84 ± 3 % of cells were plp positive. In conclusion, the results of our study showed that cells could be com pletely differentiated, from oligodendrocyte progenitors to mature, myelinating oligodendrocytes. Mature cells showed myelination of neighbor cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|