Formation of new muscle fibres and tumours after injection of cultured myogenic cells.
We examined the effects of implantation of cultured myogenic cells from a permanent cell line into soleus muscles of histocompatible adult mice. Myogenic cells (10(6) or 10(4)) were implanted into intact muscles, muscles frozen with liquid nitrogen, paralysed with botulinum toxin or reinnervated after long-term (seven months) denervation. Formation of numerous muscle fibres in myogenic cell-injected muscles raised the total number of fibres up to ten times above control by four weeks. Larger effects were found in freeze-damaged than in paralysed muscles. The new fibres had small calibers, considerable length (greater than 1.3 mm, maximum distance over which serial sections were made), were multinucleated and were oriented parallel to the large-diameter fibres of the host muscles. In some experiments beta-galactosidase, introduced into myogenic cells via retroviral transfection, was detected in small and large muscle fibres 4-20 weeks after implantation, indicating survival of the grafted cells and formation of mosaic (host-donor) and new fibres of donor origin. Muscle weight increased significantly and, rather surprisingly, a parallel increase was found in isometric tetanic tension of isolated nerve-muscle preparations; thus tension per mg muscle tissue was not different from normal. By eight weeks reduction of acetylcholine sensitivity and down-regulation of neural cell adhesion molecule to normal were observed, indicating that synaptic transmission at the new fibres was mature. After different periods of time (5-20 weeks, depending on the subclone used) tumours developed in most but not all injected limbs (37 out of 39). The tumours were destructive to the muscles and were classified as rhabdomyosarcomas. Prior to tumour formation, neural cell adhesion molecule positive cells reappeared in the muscles; since the myogenic cells initially produced differentiated muscle fibres, it appears that malignant growth is induced by factors in vivo. Thus, at present the outcome of such implantation is unpredictable.
Wernig
A
A
Physiologisches Institut, Universität Bonn, FRG.
Irintchev
A
A
Härtling
A
A
Stephan
G
G
Zimmermann
K
K
Starzinski-Powitz
A
A
D000110:Acetylcholinesterase / Q000032:analysis; D000818:Animals; D001905:Botulinum Toxins / Q000494:pharmacology; D002478:Cells, Cultured; D005615:Freezing; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D009119:Muscle Contraction; D009121:Muscle Denervation; D009132:Muscles / Q000294:innervation / Q000502:physiology / Q000637:transplantation*; D009584:Nitrogen; D010243:Paralysis; D011950:Receptors, Cholinergic / Q000032:analysis; D012190:Retroviridae / Q000235:genetics; D012208:Rhabdomyosarcoma / Q000209:etiology* / Q000473:pathology; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D011950:Receptors, Cholinergic; D000110:Acetylcholinesterase; D001616:beta-Galactosidase; D001905:Botulinum Toxins; D009584:Nitrogen