| O-2A (oligodendrocyte-type2astrocyte) progenitor cell as a bipotential cell, candifferentiate into oligodendrocyte and type2astrocyte. Seperation and purification ofO-2A progenitor cells contributes to the better understanding of their biologicalcharacteristics on proliferation and differentiation. They can also be used as a transplantedcell to cure demyelinating disease and CNS disease. However, at present, it is mainlythrough the temperature oscillation and differential adhesion method to separate and purifyO-2A progenitor cells. But in the process of collecting, some O-2A progenitor cells can notbe separated from the mixed type1astrocytes (T1As), and disappeared gradually with thegrowth and passage T1As. This experiment using erythropoietin (EPO) treating O-2Aprogenitor cells mixed in T1As, find and analyze its biological characteristics.Aims: To study the effects of EPO on proliferation of O-2A progenitor cells mixed inT1As. To observe the changes of proliferation and differentiation and other biologicalcharacteristics on EPO treated O-2A progenitor cell. To find whether it can used as celltransplantation for treatment of nervous system diseases related to cell sources provide thebasis.Methods:(1) Primarily cultured mixed glia and glial cells were isolated fromnewborn rat cortex, and then we got purified glial cells according to their difference inadhesive potential.(2) Cultured O-2A progenitor cells mixed in T1As in the mediumcontaining EPO (10U/ml) and10%NBCS, and observed the growth state of O-2Aprogenitor cells.(3) Tested the effects of proliferation of different condion supernate:[A1CM (T1As condional medium), EA1CM (EPO-treated T1As conditional medium), EACM (EPO-treated T1As mixed with O-2A progenitor cells conditional medium)] onpurified O-2A progenitor cells by MTT assays.(4) We collected the proliferating O-2Aprogenitor cells named EO-2A (erythropoetin-treated O-2A), and identified these EO-2A progenitor cells and their differentiated cells by immunofluorescent stainning method.According to the EO-2A progenitor cells in culture medium of proliferation anddifferentiation, cell growth curve was drawn and statistical analysis the pencentage ofdifferentiated cells.Results:(1) O-2A progenitor cells are highly proliferating on the layer of T1As on3-5days after the treatment by EPO (10U/ml), and they have strong dioptre.(2) A1CMhave effects of proliferation on O-2A progenitor cells, EPO, EA1CM and EACM had noeffect on the proliferation of O-2A progenitor cells.72h after the treating by EPO, theO-2A progenitor cells contacted with T1As are proliferating, but it differentiated into T2Asif without T1As contacting.(3) Compared with O-2A progenitor cells, EO-2A progenitorcells delayed the beginning of proliferation and differentiation. Immunofluorescentstaining results showed that EO-2A progenitor cells exhibited NG2and A2B5doublepositive, type-2-astrocytes (T2As) exhibited GFAP and A2B5double positive, and theirdifferentiated mature oligodendrocyte exhibited GalC and MBP double positive. EO-2Aprogenitor cells have the potentia of delaying O-2A progenitor cells’ proliferation anddifferentiation.Conclusions: The results show that EPO can make the O-2A progenitor cells mixedin T1As proliferating and this effect depends on the contact of T1As and O-2A progenitorcells. Compared with the normal cultured O-2A progenitor cells, EO-2A progenitor cellshave longer processes, and easily induced to differentiate into T2As. This may provide abasis for the cell transplantation treatment of diseases related to nervous system cells. |
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