Biogenesis Read Online Free PDF Page B

cylinder. I knew it had to be something important, but my husband never discussed his work with me, and I also assumed that Dr. Sakakibara would know, so I was the one who was surprised. […] Even after my husband disappeared, someone came once a month from Asahikawa to swap out the gas. It wasn’t such a large expense …” (Mrs. Sumiko Ishikawa).
    Mrs. Ishikawa had simply misunderstood the intent of the phone interview, but preserved winged mouse tissue became available to the world as a result. Dr. Ishikawa had stored the remainder of the culture he had requested from Mr. Miura in liquid nitrogen.
    Dr. Sakakibara and Mr. Tamura left immediately for the Ishikawa residence to speak to Mrs. Ishikawa. She consented on the spot since it would allow her husband’s work to be continued, and the tank wasbrought to the center for a culturing attempt. Dr. Ishikawa appeared to have been a very methodical person; thin slices of tissue had been deposited by organ in upwards of thirty storage tubes, and a list described the content of each.
    Yet work did not begin straight away. Normally, when frozen tissue is brought back to room temperature, much of it is dead. More accurately, upon freezing, the water content within the cells crystallizes and causes damage. With only so much tissue, it might be better, ran the long discussion, for them to let it all slumber in liquid nitrogen until regeneration technologies were somewhat more advanced, just as Dr. Ishikawa seemed to have willed.
    Dr. Akedera’s position was that they should culture all of the tissues and re-preserve them under better conditions, whereas Dr. Sakakibara emphasized the dangers of wasting the material. Messrs. Tamura and Miura were also in favor of storing the tissues, so there was something of a standoff. In the end, it became evident that epidermal tissue had been parceled among the various storage tubes,and an agreement was made to defrost one. Hence both Dr. Akedera and Dr. Sakakibara managed to stick to their guns: analysis and preservation, respectively.
    Of all tissues, the epidermis includes cells that are the easiest to culture, with the highest success rates. Rapidly defrosted in a 37°C incubator, the tissue was cut into small pieces in a Petri dish, with some reserved for extracting nucleic acid and the rest placed in deactivated serum. After twenty-fours had passed, the material was transferred to a medium so that culturing could continue. According to the records, Dr. Akedera performed all of this work himself and alone with the utmost care.
    Cells seeping beyond the tissue fragment were observed ten hours into the culturing process, and comparable growth obtained in all the bases after twenty-four more hours. At this point tissue fragments were retrieved from the Petri dish to be re-frozen.
    Meanwhile, one gram of tissue fragment was placedin liquid nitrogen and destroyed, and the nucleic acids extracted using the MAD method 8 –– which is known to damage genomic material (DNA) the least – and stored at −70°C in a state of ethanol precipitate.
    Seventy-two hours into the process, overgrown cells were carefully detached from the Petri dish and replanted using EDTA isolation (later, it was imagined that this step was to blame for the failed culturing). A portion of the detached cells went into serum mixed with DMSO and stored in liquid nitrogen.
    The experiment was proceeding smoothly, and Dr. Akedera seemed to have achieved his initial goal of preservation at the cellular and genetic level. Rapid changes began to occur ninety-six hours into culturing, however.
    “Although they weren’t confluent [covering the surface without gaps], on the morning of the fourth day, the cells in the nearly thirty Petri dishes had floated up all at once. I ran to tell Dr. Akedera, and then things went a little haywire,”recalled Mr. Miura, who was serving as an assistant at that point. “The culturing work was being done with great care with varying