The donor somatic cell karyoplast
is one of the important factors affecting the efficiency of somatic cell animal cloning.
The battle to prevent the transmission of mitochondrial DNA disease: is karyoplast transfer the answer?
Metaphase II karyoplast transfer from human in-vitro matured oocytes to enucleated mature oocytes.
GJ: In ons laboratorium is gebruik gemaak van morula blastomere, weens hul ontwikkelingsplastisiteit, as donor karioplast.GJ (translation): In our laboratory we used morula blastomeres, because of their differentiating plasticity, as donor karyoplast
(57) Researchers then used a karyoplast
fusion technique, using an extract from the Sendai virus, (58) which produced reconstructed oocytes which maintained the spindle-chromosomal complexes transferred from the mature oocyte.
To confirm the enucleation of oocyte, the aspirated karyoplast
in the pipette was visualized under fluorescent light.
Green fluorescent protein (GFP) gene was used as a foreign gene and a non-transfected somatic cell was utilized as a control karyoplast
. Monolayers of porcine cells were established and subsequently transfected with a GFP-expressing gene (pEGFP-N1) using three types of transfection reagents (LipofectAMINE PLUS, FuGENE 6 or ExGen500).
Our earlier studies using asbestos fiber--treated cytoplasts and fusion with non-treated karyoplasts
provided circumstantial evidence of the role of mitochondria in fiber genotoxicity (Xu et al.
from multinucleated fibroblast cells contain centrosomes and change their morphology to mi totic cells.
The resulting pronuclei (karyoplasts
) in the donor oocytes were microsurgically removed and discarded, and the pronuclei from the patient's oocytes were transferred into the donor's enucleated cells (cytoplasts).
The aspirated karyoplasts
were exposed to ultraviolet-light to confirm the presence of nuclei.