Ovary biotechnology

Several advances in ovarian biotechnology may accelerate ovarian stimulation research. 

For more information: 2017's hottest biotechnology is focused on ovaries!

Digital images and flow diagrams illustrating the pump pathways of the (a) Solo-MFP, (b) Duet-MFP and (c) Quintet-MFP systems. (d) Illustration of the pump mechanism of the electromagnetic Quintet-MFP. AC, acceptor module; DO, donor module; T, tissue module.

Multiple follicle culture in the Solo-MFP (stages i–iii); microfluidic culture supported follicle growth from primary stage to antral stage (iv–vi); and, following hCG stimulation, completion of the first meiotic division by the oocyte was achieved oocyte completed the first meiosis indicated by well-organized microtubule fibres (green), tightly aligned chromosomes (blue), and the round appearance of F-actin (red; stages vii–viii). (b) Granulosa cells showed similar morphological changes as seen in vivo following luteinization, indicated by hypertrophy and decreased nucleus to cytoplasm ratio. (c,d) Ovarian hormone secretions of oestradiol (E2) and progesterone (P4; c) and inhibin A and inhibin B (d) over 28 days of culture in the Solo-MFP. (e,f) Comparison of E2 (e) and P4 (f) secretion rates between microfluidic and static cultures. Graphs in b,fdisplay average+s.d. *P<0.05 comparison of the number of nuclei per mm3 in follicles before and after hCG treatment (b), and hormone secretion rates between microfluidic and static cultures (e,f). Scale bar, 50 μm (a) and 10 μm (b). n=3–6 replicates for both the microfluidic and static culture.

(a) Ectocervix histology, and Ki67 and PR staining at the end of follicular (day 0) and luteal (day 14) phases. (b) Histology of liver microtissues before (day −14) and after (day 14) hCG treatment. (c) Human albumin production over 28 days of microfluidic culture. (d) Production of IL8 and VEGF over 28 days of microfluidic culture. (e) Histology of ovarian tissue in the Quintet-MFP on day 0 (pre hCG) and day 8 (continuously cultured with hCG). (f) Ovarian progesterone secretion with and without hCG treatment. Graphs in c,d,f display average+s.d.’. Scale bar, 10 μm (a), 25 μm (b), and 100 μm (e). CL, corpus luteum; IL8, interleukin 8; VEGF-A vascular endothelial growth factor A. n=3 replicates of the integrated tissue culture in the Quintet-MFP.

a) Light microscopy image of 3βHSD expression (purple) along edge of follicle cultured in 30° scaffold after 4 days of culture. (b) Estradiol secretion in media of follicles cultured in 3D printed scaffolds collected at day 2 and day 8 of culture. (c,d) MII egg with extruded polar body was released from a follicle cultured in 60° scaffold and contains condensed chromatin (blue) along the spindle (green). (e,f) Scanning electron micrograph demonstrating follicle (arrows) wedged underneath three layers of 60° scaffold struts (identified as layers A, B, C) and cultured for 2 days. Scale bars: (c) 50 μm; (a,e) 100 μm; (d,f) 10 μm.

(a) Whole-mount fluorescent image of GFP+ follicles (green) implanted within the ovarian bursa of GFP− mice removed 8 weeks after surgery. (b,c) Vascularization shown in representative images of immunostaining for endothelial marker platelet endothelial cell adhesion molecule (PECAM) (red) or pericyte marker PDGFRβ1 (green) expression in corpus luteum, antral follicles and interstitial space of bioprosthetic ovary removed 8–10 weeks post-surgery. DNA counterstained blue. (d) Quantification of vessels containing red blood cells within ovarian bioprosthesis collected 1 or 3 weeks post-surgery. (e,f) H&E stained cross-sections of ovarian bioprostheses, removed 3 weeks after surgery, contained vessels and primordial, primary, secondary and antral follicles within the gelatin scaffold struts. (g) Serum analysis for peptide hormones anti-Müllerian hormone (AMH) and inhibin A detected within ovariectomized animals with empty scaffold implant (OVX+Sham) or with bioprosthetic ovary (OVX+Implant). (h) GFP+ pup (black arrow) born to GFP− bioprosthesis recipient female. (i) GFP+ pup born from bioprosthesis sired liters with GFP− CD1 female to create a mixed litter of GFP+ and GFP− pups (grand-pups of ovary recipient). §, scaffold strut. +, vessels. Scale bars: (a) 1 mm; (b,c,e,f) 50 μm. All data are presented as mean±s.e.m. ND, not detected.