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I.R.M., New York University Medical Center, New York, and the Medical Neurology Branch, National Institute of Neurological Diseases and Stroke, Bethesda, Maryland.
With our new "explant-reexplanting" technique, abundant growth of mature human muscle in long-term tissue culture was achieved, and with the "sandwich" technique several histochemical reactions were obtained on serial cross sections of the cultured fibers. An advanced degree of maturation but lack of differentiation into reciprocally staining fiber-types was demonstrated. For electron-microscopic and electronmicroscopic-histochemical study, a method was developed in which the embedded fibers of greatest potential interest were identified by light microscopy and punched out by our specially-designed hollow drill. This selection procedure is critically important when the goal is to study in cultured diseased human muscle: (1) successive stages of development and (2) certain structural changes that often occur only in some fibers and only in certain regions of those fibers. The electronmicroscopic-histochemical appearance of developing cultured muscle fibers correlated well with the fresh-frozen light microscopic histochemical cross-sections and longitudinal whole preparations of similar fibers.
This study was supported by the Muscular Dystrophy Associations of America (Dr. Askanas).
Received for publication July 11, 1974.
Dr. Askanas address is New York University Medical Center, 400 E. 34th St., New York, NY 10016.
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