Gene transfer into satellite cell from regenerating muscle: bupivacaine allows beta-Gal transfection and expression in vitro and in vivo.
A large bulk of experimental evidence (15) suggests that myogenic cell transfer can be regarded as a promising therapeutic approach in the cure of inherited pathologies. In particular, it has been shown that primary myoblasts obtained from embryonic or neonatal muscles allows the recovery of the normal phenotype in defective muscle tissues. The utilization of this approach in clinical settings still bears heavy limitations. Apart from the legal and ethical difficulties, the use of muscles obtained from aborted fetus is challenged by a large risk of rejection, due to the incompatibility between donor and recipient. In this context based on the genetic alteration and reimplanting of the patient's own satellite cells, appears an approach attractive. Myoblasts derived from satellite cells are the obligate candidates for experiments, but the production of sufficient cell numbers is a major problem. Local anesthetics [Bupivacaine (1-n-butyl-DL-piperidine-2-carboxylic acid-2, 6-dimethyl anilide hydrochloride) and related molecules] had been used to induce myofiber damage (and thus satellite cells proliferation) and thereby may represent a tool for increasing the yield of myoblasts from adult muscles (1,9,17). We will show that satellite cells obtained from adult muscles after bupivacaine injection can be transfected in vitro and that the transfected gene is expressed in vitro and in vivo, after reimplantation of the modified myoblasts in recipient muscles.
Department of Experimental Biomedical Sciences, University of Padua, Italy.
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M
Cantini
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Massimino
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C
Catani
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Rizzuto
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Brini
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Carraro
D000818:Animals; D002045:Bupivacaine / Q000494:pharmacology*; D002478:Cells, Cultured; D015870:Gene Expression; D018014:Gene Transfer Techniques; D009132:Muscles / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D051381:Rats; D017208:Rats, Wistar; D012038:Regeneration; D001616:beta-Galactosidase / Q000235:genetics*
D001616:beta-Galactosidase; D002045:Bupivacaine