Evaluation of the meiotic spindle apparatus in metaphase II human oocytes following cytoplasmic donation.
OBJECTIVE
To determine if the removal of cytoplasm from metaphase II human donor oocytes damages the meiotic spindle apparatus.


METHODS
Cryopreservation of metaphase II human oocytes was performed using a fast-freeze, fast-thaw protocol. Upon thaw, oocytes were incubated for 3-4 h and then used for cytoplasmic donation (test oocytes). Oocytes thawed but not used for donation served as controls. Test and control oocytes were fixed using a microtubule-stabilizing buffer. Tubulin was localized using antitubulin monoclonal antibody. Chromosomes were identified by counterstaining with DAPI.


RESULTS
Forty-four oocytes had cytoplasm removed (test group) while 12 were not used for the procedure (controls). Twenty-three oocytes survived the donation procedure. Rates of normal spindle structure for the control and test groups were 21/23 (91.3%) and 12/12 (100%), respectively.


CONCLUSIONS
The removal of cytoplasm from a metaphase II human donor oocyte does not appear to significantly increase the damage to chromosome alignment or to the spindle structure.
Jones
E L
EL
Department of Obstetrics/Gynaecology, Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, 601 Colley Avenue Norfolk, Virginia 23507, USA.
Boyd
C A
CA
Dowling-Lacey
D
D
Wright
D
D
Mayer
J F
JF
Lanzendorf
S E
SE
D000328:Adult; D015925:Cryopreservation; D003593:Cytoplasm / Q000637:transplantation*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008540:Meiosis / Q000502:physiology*; D008677:Metaphase; D018587:Oocyte Donation; D009865:Oocytes / Q000166:cytology*; D008941:Spindle Apparatus