. 2021 Dec 28;106(2):291–315. doi: 10.1093/biolre/ioab242

Table 4.

Maturation and morphology of oocytes derived from cultured follicles in humans

Patient	Age	Initial follicle stage and isolation method	Culture system and interval	Basal media, supplement and O₂ level	Oocyte assessment and outcomes	References
Cancer, aplastic anemia, beta-thalassemia	5–33 years	Secondary; Mechanical	1. 0.5% (w/v) alginate-embedded individual follicle culture till follicle diameter = 400–500 μm with antrum formation; 2. Individual follicle culture in low attachment plate till 30–40 days; 3. Individual follicle incubation with 1.5 IU/ml hCG and 10 ng/ml EGF for 16 h	1/2. αMEM Glutamax and F-12 Glutamax at 1:1, 5 μg/ml insulin, 10 mIU/ml rhFSH; 3. αMEM, 10% FBS, 10 mIU/ml rhFSH; 1–3: no O₂ control	4/20 MIIs with typical spindle structure by immunofluorescence, polar body fragmentation by light microscopy	[69]
Elective cesarean section	25–39 years	Unilaminar (primordial, intermediary or primary); Mechanical	1. Ovarian tissue culture for 8 days; 2. Secondary follicle individual culture in 96-well V-bottom plate for 8 days; 3. COC group culture on track-etched nucleopore membrane for 4–6 days; 4. 24 h of IVM	1. McCoy’s 5A, 10 ng/ml insulin, 50 μg/ml ascorbic acid, 1 ng/ml rhFSH; 2/3. McCoy’s 5A, 10 ng/ml insulin, 50 μg/ml ascorbic acid, 1 ng/ml rhFSH, 100 ng/ml rhACVA; 4. SAGE IVM media, 75 mIU/ml rhFSH, 75 mIU/ml rhLH; 1–4: no O₂ control	9/32 MIIs with typical spindle structure by immunofluorescence, large polar body by light microscopy	[71]
Benign or malignant gynecologic conditions	22–45 years	Unilaminar (primordial or intermediary); Mechanical	1. Ovarian tissue culture for 3 weeks; 2. Secondary follicle group culture in 96-well round-bottom ultra-low attachment plate for 6 weeks; 3. 48 h of IVM	1. αMEM, 5 μg/ml insulin, 15 mIU/ml rhFSH; 2. αMEM, 5 μg/ml insulin, 15 mIU/ml rhFSH, 100 ng/ml rhAMH in week 0–3 and 100 ng/ml AMH-Ab in week 4–6; 3. SAGE IVM media, 75 mIU/ml rhFSH, 75 mIU/ml rhLH; 1/2: no O₂ control; 3: 5% O₂	3/14 MIIs with normal spindle/polar body size/position and typical ultrastructure by polarized microscopy and electron microscopy	[73]

Patient

Age

Initial follicle stage and isolation method

Culture system and interval

Basal media, supplement and O₂ level

Oocyte assessment and outcomes

References

Cancer, aplastic anemia, beta-thalassemia

5–33 years

Secondary;
Mechanical

1. 0.5% (w/v) alginate-embedded individual follicle culture till follicle diameter = 400–500 μm with antrum formation;

2. Individual follicle culture in low attachment plate till 30–40 days;

3. Individual follicle incubation with 1.5 IU/ml hCG and 10 ng/ml EGF for 16 h

1/2. αMEM Glutamax and F-12 Glutamax at 1:1, 5 μg/ml insulin, 10 mIU/ml rhFSH;
3. αMEM, 10% FBS, 10 mIU/ml rhFSH;
1–3: no O₂ control

4/20 MIIs with typical spindle structure by immunofluorescence, polar body fragmentation by light microscopy

[69]

Elective cesarean section

25–39 years

Unilaminar (primordial, intermediary or primary);
Mechanical

1. Ovarian tissue culture for 8 days;

2. Secondary follicle individual culture in 96-well V-bottom plate for 8 days;

3. COC group culture on track-etched nucleopore membrane for 4–6 days;

4. 24 h of IVM

1. McCoy’s 5A, 10 ng/ml insulin, 50 μg/ml ascorbic acid, 1 ng/ml rhFSH;
2/3. McCoy’s 5A, 10 ng/ml insulin, 50 μg/ml ascorbic acid, 1 ng/ml rhFSH, 100 ng/ml rhACVA;
4. SAGE IVM media, 75 mIU/ml rhFSH, 75 mIU/ml rhLH;
1–4: no O₂ control

9/32 MIIs with typical spindle structure by immunofluorescence, large polar body by light microscopy

[71]

Benign or malignant gynecologic conditions

22–45 years

Unilaminar (primordial or intermediary);
Mechanical

1. Ovarian tissue culture for 3 weeks;

2. Secondary follicle group culture in 96-well round-bottom ultra-low attachment plate for 6 weeks;

3. 48 h of IVM

1. αMEM, 5 μg/ml insulin, 15 mIU/ml rhFSH;

2. αMEM, 5 μg/ml insulin, 15 mIU/ml rhFSH, 100 ng/ml rhAMH in week 0–3 and 100 ng/ml AMH-Ab in week 4–6;

3. SAGE IVM media, 75 mIU/ml rhFSH, 75 mIU/ml rhLH;

1/2: no O₂ control; 3: 5% O₂

3/14 MIIs with normal spindle/polar body size/position and typical ultrastructure by polarized microscopy and electron microscopy

[73]

AMH, anti-Müllerian hormone; AMH-Ab, neutralizing anti-human anti-Müllerian hormone antibody; COC, cumulus–oocyte complex; EGF, epidermal growth factor; FBS, fetal bovine serum; hCG, human chorionic gonadotropin; IVM, in vitro maturation; αMEM, alpha minimum essential medium; MII, metaphase II; rhACVA, human recombinant activin A; rhAMH, recombinant human anti-Müllerian hormone; rhFSH, recombinant human follicle-stimulating hormone; rhLH, recombinant human luteinizing hormone.