Improved Method for Establishment of Paroxysmal Nocturnal Hemoglobinuria Immortalizing Cell Lines by EB Virus Transfection

The establishment of paroxysmal nocturnal hemoglobinuria immortalizing cell lines by Epstein-Barr virus transfection is more difficult and takes more time than that of normal cell lines. Adding a kind of antioxidant (AA2) in the culture system was performed. Immortalizing cell lines were successfully established from each of the two cases thus treated, and the time required became shorter, while only one cell line was acquired from the two cases without addition of AA2 during culture. This result suggested that oxidative damage was possibly the reason of the difficulty to bread the cell lines; adding antioxidant in culture system helped to increase the success rate and to shorten the time of establishing the immortalizing cell lines. By adding antioxidant, the percentage of CD59- cell had almost no reduction after the cell line establishment process. However, we had only performed this trial in two cases and more experiments need to be done before the effect of antioxidant can be finally confirmed .