Comparison of different electrofusion treatments to be used in rabbit somatic cloning

• Autores: María José Escribá Pérez, Miguel Angel Silvestre Camps, A. M. Saeed, R.P Cervera, Fernando García Ximénez
• Localización: Investigación agraria. Producción y sanidad animales, ISSN 0213-5035, Vol. 17, Nº 1-2, 2002, págs. 49-58
• Idioma: inglés
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• Resumen

  • The present study examined the efficiency of different electrofusion treatments (A, B and C), in terms of fusion and lysis rates when fibroblasts or cumulus cells were used as nuclear donors. Regardless of nuclear donor cell used, treatment B (a pulse of 2.8 kVcm' for 60 usec, applied three times 30 min apart) gave acceptable fusion rates (fibroblasts: 59 %; cumulus cells: 69-95 %) with low lysis rates (fibroblats: 1.6 %; cumulas cello:

    o %). Additionally, treatment A (two pulses of 1.5 kVcmt for 60 usec, applied three times 30 min apart) presented a high disconnection rate (39 %), while treatment C (three pulses of 3.6 kVcm' for 20 .usec, repeated once 90 min apart) supposed an excessively high lysis rate (19-22 %) Hypo-osmolarity in the B-pulsing medium reduces fusiun rate (0.25 M: 81 % vs. 0.3 M: 95 % P < 0.05), as well as cleavage rate (0.25 M: 23 %s. 0.3 M: 70 %, P <.0.05). Thus, the isosmolar treatment B is an efficient electrofusion treatment in rabbit somatic cloning when these cellular types are used as nuclear donors. On the other hand, the pulse order in which the fusion occurs determines differences on further cleavage rates.