Table 2.
Maturation and competence of oocytes derived from cultured follicles in domestic animals
| Species | Age | Initial follicle stage and isolation method | Culture system and interval | Basal media, supplement and O2 level | Oocyte assessment and outcomes | References |
|---|---|---|---|---|---|---|
| Cattle | Unknown | Early antral 0.5–0.7 mm Mechanical |
1. COCs with partial presence of granulosa cells embedded in collagen gels, 4–6 per gel, 14 days until antral follicle-like structure formed 2. COCs extracted mechanically and cultured with additional cumulus cells and inseminated 14 days |
1–2. TCM199, 10% FCS, 4 mM hypoxanthine, 5% CO2 in air | 6/22 (27%) MII 10/24 (42%) fertilization 21/135 (18%) 2-cell 6/135 (4%) blastocyst 3 blastocysts/3 recipients 1 live offspring | [41] |
| Cattle | Unknown | Early antral 0.4–0.7 mm Mechanical |
1. COCs with partial presence of granulosa cells, 3–4 placed onto collagen-coated membrane inserts 2. COCs with partial presence of granulosa cells individually cultured in collagen-coated 96-well plates, 14 days until antral follicle-like structure formed 3. COCs removed mechanically, 6–10 per drop maturation media 14 days |
1–2. TCM199, 5% FBS, 4 mM hypoxanthine, 0.1 mg/ml estradiol, 4% PVP (MW 360 K) 3. TCM199, 5% FBS, 0.1 mg/ml sodium pyruvate, 20 mIU/ml rbFSH 5% CO2 in air |
51/58 (88%) MII 48/51 (94%) fertilization 20/48 (27%) cleavage 7/48 (17%) blastocyst 10/123 (8%) blastocyst per total number oocytes recovered 4 blastocysts/4 recipients 1 live offspring | [42] |
| Goat | Adult, 1–3 year-old | Preantral (secondary) ≥ 150 μm Mechanical |
1. Individual culture in drops on Petri dish 2. COCs removed for IVM/IVF 18 days |
1. α-MEM, 3 mg/ml BSA, 1 mg/ml fetuin, 10 μg/ml insulin, 2 mM glutamine, 2 mM hypoxanthine, 50–100 ng/ml rbLH, 50–100 ng/ml EGF, 100 ng/ml rbFSH Days 0–6, 500 ng/ml rbFSH Days 6–12, 1000 ng/ml rbFSH Days 12–18 2. TCM199, 1 mg/ml BSA, 22 μg/ml pyruvate, 100 μM cysteamine, 1 μg/ml estradiol, 0.5 μg/ml rbFSH, 5 μg/ml rbLH, 10 ng/ml EGF, 50 ng/ml IGF1 5% CO2 in air |
2/32 (6%) MII One 8-cell One 16-cell |
[43] |
| Goat | Adult, 1–3 year-old | Preantral (secondary) follicles, 200 μm Mechanical |
1. Individual culture in drops on Petri dish 2. COCs removed for IVM/IVF 18 days |
1. α-MEM, 3 mg/ml BSA, 1 mg/ml fetuin, 10 μg/ml insulin, 2 mM glutamine, 2 mM hypoxanthine, 50 ng/ml GH, 100 ng/ml rbFSH Days 0–6, 500 ng/ml rbFSH Days 6–12, 1000 ng/ml rbFSH Days 12–18 2. TCM199, 1 mg/ml BSA, 22 μg/ml pyruvate, 100 μM cysteamine, 1 μg/ml estradiol, 0.5 μg/ml rbFSH, 5 μg/ml rbLH, 10 ng/ml EGF, 50 ng/ml IGF1 5% CO2 in air |
10/20 (50%) MII 1/35 (3%) 2-cell embryo, progressed to compact morula |
[44] |
| Goat | Adult and prepubertal | Secondary Mechanical |
Single follicle 2D 18 days |
α-MEM, 3 mg/ml BSA, 1 mg/ml fetuin, 10 μg/ml insulin, 2 mM glutamine, 2 mM hypoxanthine, 100 ng/ml VEGF, 100 ng/ml rbFSH Days 0–6, 500 ng/ml rbFSH Days 6–12, 1000 ng/ml rbFSH Days 12–18 5% CO2 in air |
2D: four 2-cell embryos 3D: one 8-cell embryo |
[45] |
| Goat | Adult, 1–3 year-old | Multi-layer secondary follicles (≥200 μm) Mechanical |
1. Individual follicle embedded in 0.5% (w/v) alginate, 48-well plates, 18 days 2. Group of 5 follicles embedded in 0.5% (w/v) alginate, fibrin-alginate or hyaluronate, 18 days 3. 5 follicles/bead in fibrin-alginate, 30 days 4. COCs removed mechanically for maturation |
1–3. α-MEM, 3 mg/ml BSA, 10 ng/ml insulin, 2 mM glutamine, 2 mM hypoxanthine, 0.9 mM pyruvate, 100 ng/ml rbFSH Days 0–6, 500 ng/ml rbFSH Days 6–12, 1000 ng/ml rbFSH Days 12–18 2–3. same as 1 with 100 ng/ml VEGF 4. TCM199, 0.4% BSA, 5 mM ionomycin, 1 mM 60DMAP, 10% FCS 5% CO2 in air |
1. Single follicle in Alginate 2/14 (14%) MII 2. 5 follicles/bead Alginate: no oocytes matured Fibrin-alginate: 5/17 (29%) MII Hyaluronate: none matured 3. Fibrin alginate 30 days 7/12 (58%) MII 3/4 parthenotes cleaved and developed to 8-cell |
[46] |
| Goat | Adult, 1–3 year-old | Early antral (350 μm) Mechanical |
1. Individual culture in drops on Petri dish 2. COCs removed for IVM/IVF 18 days |
1. α-MEM, 3 mg/ml BSA, 10 μg/ml insulin, 2 mM glutamine, 2 mM hypoxanthine, 0.9 mM pyruvate, 50 ng/ml GH, 300 μg/ml anethole 2. TCM199, 1 mg/ml BSA, 1 mM pyruvate, 10 μM cysteamine, 1 μg/ml estradiol, 0.5 μg/ml rbFSH, 5 μg/ml rbLH, 10 ng/ml EGF, 50 ng/ml IGF1 5% CO2 in air |
56/97 (58%) MII 29/79 (37%) cleavage 10/77 (13%) parthenotes 14/60 (23%) cleavage IVF 6/60 (10%) embryos IVF 6 cleaved embryos/3 recipients 1 pregnancy to Day 30 gestation |
[47] |
| Sheep | Preantral follicles 250–400 μm Mechanical |
1. Microdrops or agar gel 6 days 2. COCs removed for IVM/IVF |
1. TCM 199, serum-free, 2 μg/ml T4, 2 μg/ml ovine FSH, insulin (2.5–15 ng/ml), IGF1 (5–20 ng/ml), GH (0.5–2 mIU/ml), TGF-β (1–15 ng/ml) 2. 10 μg/mL each of ovine FSH, Ovine LH, 1 mg/ml BSA, 1 μg/mL estradiol 5% CO2 in air |
T4+ FSH + ITS+IGF1 + GH 159/250 (64%) MII 35/159 (22%) 2-cell 3/36 (1%) morula T4 + FSH + IGF1 + GH 170/250 (68%) MII 43/170 (26%) 2-cell 7/43 (1%) morula |
[48] | |
| Sheep | Prepubertal | Preantral with evident theca layer (secondary) 170 ± 30 μm Early antral Mechanical |
Individual follicles placed in 96-V-well plates 14 days |
1. α-MEM, 2% FCS, 1% insulin, 1 μg/ml ovine FSH 5% CO2 in air |
Preantral – no data Early Antral: 47/110 (43%) MII Parthenotes 51/150 (34%) <16-cell 8/150 (5%) >16-cell IVM/IVF 107/336 (32%) fertilized 97/107 (91%) <16-cell 11/107 (10%) >16-cell |
[49] |
| Sheep | Adult (1–3 years) |
Preantral (secondary) >200 μm Mechanical |
Individual follicles in drops in Petri dishes 18 days |
α-MEM, 3 mg/ml BSA, 10 μg/ml insulin, 2 mM glutamine, 2 mM hypoxanthine, 50 ng/ml IGF1 or LIF or KL, 100 ng/ml ovine FSH Days 0–6, 1000 ng/ml ovine FSH Days 6–12, 200 ng/ml ovine FSH Days 12–18 5% CO2 in air |
50 ng/ml LIF 8/27 (29%) MII 7/12 (58%) 8-cell parthenotes 50 ng/ml KL 32% fertilization 1 morula after IVF |
[50, 52] |
| Sheep | Prepubertal to 10 month-old | Preantral 160–240 μm 0.1% Collagenase IA and 0.01% DNase I, followed by Mechanical |
1. Individual follicles in drops in Petri dishes 2. COCs removed mechanically for maturation 20 days |
1. α-MEM, 1.25 mg/ml BSA, 6.25 μg/mL insulin, 2 mM glutamine, 2 mM hypoxanthine,100 ng/ml ovine FSH alone or with 50 ng/mL LIF 2. TCM199, 10 ng/ml EGF,100 μM cysteamine |
LIF + ovine FSH 56% MII |
[51] |
| Pig | Prepubertal | Preantral (multi-layer) 200–310 μm; 296 ± 9 μm Mechanical |
1. 24-well dishes, 3 follicles per well 2. COCs removed mechanically for maturation and IVF 4 days |
1. NCSU23, 7.5% porcine serum, 3.5 μg/ml insulin, 1.5 ng/ml ovine FSH 2.NCSU23, 0.12 μg/ml ovine FSH, 2.5 μg/ml ovine LH, 20 ng/ml EGF 5% CO2 in air |
IVF 124/244 (51%) MII 60/114 (53%) fertilization 26/60 (43%) 2-cell 8/60 (13%) blastocyst ICSI 57/89 (64%) fertilization 29/57 (51%) 2-cell 12/57 (21%) blastocyst |
[53, 54] |
| Buffalo | Mature | Preantral, (multi-layer) 265–295 μm 1% trypsin, then mechanical |
1. 2–3 follicles per well; supplemented with ovarian mesenchymal cells or cumulus cells 2. COCs removed mechanically for maturation and IVF 100 days |
1. α-MEM, 10% steer serum, 6.25 μg/mL insulin, 0.23 mM pyruvate, 2 mM glutamine, 2 mM hypoxanthine, 3 μg/ml rbFSH 2. TCM199, 10% steer serum, 0.05 IU/ml rbFSH 5% CO2 in air |
82–100% oocyte recovery rate 38–46% cleavage 11–12% morulae/blastocyst |
[55] |
2D, 2-dimensional; 3D, 3-dimensional; BSA, bovine serum albumin; CO2, carbon dioxide; COC, cumulus–oocyte complex; DNase, deoxyribonuclease; DMAP, dimethylaminopyridine; EGF, epidermal growth factor; FBS, fetal bovine serum; FCS, fetal calf serum; GH, growth hormone; ICSI, intracytoplasmic sperm injection; IGF1, insulin-like growth factor 1; ITS, insulin–transferrin–selenium; IVF, in vitro fertilization; IVM, in vitro maturation; KL, kit ligand; LIF, leukemia inhibitory factor; αMEM, alpha minimum essential medium; MII, metaphase II; MW, molecular weight; NCSU, North Carolina State University; O2, oxygen; PVP, polyvinylpyrrolidone; rb-FSH, recombinant bovine follicle stimulating hormone; rbLH, recombinant bovine luteinizing hormone; T4, thyroxine; TCM199, tissue culture media 199; TGF-β, transforming growth factor-β; VEGF, vascular endothelial growth factor.