TY - JOUR
T1 - Technical aspects of the piezo, laser-assisted, and conventional methods for nuclear transfer of mouse oocytes and their efficiency and efficacy
T2 - Piezo minimizes damage of the ooplasmic membrane at injection
AU - Chen, Shee Uan
AU - Chao, Kuang Han
AU - Chang, Chia Y.I.
AU - Hsieh, Fon Jou
AU - Ho, Hong Nerng
AU - Yang, Yu Shih
PY - 2004/4/1
Y1 - 2004/4/1
N2 - Assessment of the advantages and disadvantages of the piezo, laser, and conventional methods for nuclear transfer has remained elusive. Furthermore, although the piezo method had been used by some investigators for research of sperm injection and nuclear transfer for several years, many researchers have failed to operate the technique smoothly and achieve reproducible results. The procedures of nuclear transfer using piezo were ascertained and described in detail. Mouse oocytes were enucleated, and injected with cumulus cells using the piezo, laser, or conventional methods. We investigated the time needed and survival of nuclear transfer. Development was compared among the three methods and parthenogenetic control specimens. The average time of nuclear transfer for each oocyte was significantly shorter using the piezo (118 ± 9 s) and laser methods (120 ± 11 s) than using the conventional method (170 ± 11 s). The damage rate was smaller for the piezo group (10%) than the laser (37%) and conventional (40%) groups. The percentages of blastocyst formation (14%, 12%, and 11%) and the number of nuclei of blastocysts (54 ± 13, 51 ± 11, and 52 ± 12) were similar among the piezo, laser, and conventional groups, but significantly lower than for the control group (83%, 105 ± 14). The piezo technique is more efficient than the conventional method for nuclear transfer, The laser method is easy to operate, but the equipment is expensive. In addition, piezo induced fewer traumas while breaking the membrane than the aspiration techniques used in the laser and conventional methods.
AB - Assessment of the advantages and disadvantages of the piezo, laser, and conventional methods for nuclear transfer has remained elusive. Furthermore, although the piezo method had been used by some investigators for research of sperm injection and nuclear transfer for several years, many researchers have failed to operate the technique smoothly and achieve reproducible results. The procedures of nuclear transfer using piezo were ascertained and described in detail. Mouse oocytes were enucleated, and injected with cumulus cells using the piezo, laser, or conventional methods. We investigated the time needed and survival of nuclear transfer. Development was compared among the three methods and parthenogenetic control specimens. The average time of nuclear transfer for each oocyte was significantly shorter using the piezo (118 ± 9 s) and laser methods (120 ± 11 s) than using the conventional method (170 ± 11 s). The damage rate was smaller for the piezo group (10%) than the laser (37%) and conventional (40%) groups. The percentages of blastocyst formation (14%, 12%, and 11%) and the number of nuclei of blastocysts (54 ± 13, 51 ± 11, and 52 ± 12) were similar among the piezo, laser, and conventional groups, but significantly lower than for the control group (83%, 105 ± 14). The piezo technique is more efficient than the conventional method for nuclear transfer, The laser method is easy to operate, but the equipment is expensive. In addition, piezo induced fewer traumas while breaking the membrane than the aspiration techniques used in the laser and conventional methods.
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U2 - 10.1002/jez.a.20037
DO - 10.1002/jez.a.20037
M3 - Article
C2 - 15039993
AN - SCOPUS:1942488984
SN - 0022-104X
VL - 301
SP - 344
EP - 351
JO - Journal of Experimental Zoology Part A: Comparative Experimental Biology
JF - Journal of Experimental Zoology Part A: Comparative Experimental Biology
IS - 4
ER -