Table 1. Chronological advances in in vitro culture of bovine preantral follicles*.
| References/Base medium | Follicle Category/Culture system | Tested compounds/Main findings |
| McLaughlin and Telfer, 2010 - McCoy´s 5a Bicarbonate (20mM Hepes, 3mM GLUT, 0.1% BSA, 2.5 µg/ml TRAN, 4 ng/ml SEL, 10 ng/ml INS and 50 µg/ml AA) | Primordial - In situ (6 days) Secondary - Isolated 2D (15 days) | 100 ng/ml Act A or 50 ng/ml FSH - Primordial follicle - ↑ activation and growth; Act A - ↑ follicle and oocyte growth and antrum formation |
| Tang et al., 2012 - α-MEM (100 ng/ml FSH, 1.25 mg/ml BSA, 1 µg/ml INS, 5.5 µg/ml TRAN and 5 ng/ml SEL) | Preantral - In situ (22 days) | 200 ng/ml GDF-9 or 100 ng/ml bFGF - GDF-9+FSH and bFGF+FSH - ↓ apoptosis and ↑ activation; GDF9+FSH - ↑ follicle diameter |
| Rossetto et al., 2012 - Base media (20mM Hepes, 3mM GLUT, 0.1% BSA, 2.5 µg/ml TRAN, 4 ng/ml SEL, 10 ng/ml INS, 50 µg/ml AA, 100 ng/ml Act A and 100 ng/ml FSH) | Secondary - Isolated 2D (16 days) | Different base media: αMEM, McCoy and TCM199 - TCM199 - ↑ follicle viability, diameter and antrum formation |
| Passos et al., 2013 - α-MEM (10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 3 mg/ml BSA, 2mM GLUT, 2mM HYP, 50 µg/ml AA) | Secondary - Isolated 2D (12 days) | 50 ng/ml BMP-15 and 50 ng/ml FSH - BMP-15 and FSH alone - ↑follicular volume and antrum formation; BMP-15+FSH - ↑ atresia |
| Sun and Li, 2013 - α-MEM (0.23 mM PYR, 1.5 mM GLUT, 2 mM HYP, 7.5% FBS, 5 µg/ml INS, 5 µg/ml TRAN, 5 ng/ml SEL) | Primary - Isolated 3D (Type I collagen - 21 days) | 0.25 µg/ml FSH, 5 IU/ml LH, 0.5 µg/ml E2, 25 ng/ml EGF and 50 ng/ml bFGF - All factors - ↑ follicle diameter and antrum formation |
| Silva et al., 2014 - α-MEM (3 mg/mL BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYP, 50 µg/ml AA) | Secondary - Isolated 2D (18 days) | 100 ng/ml Act A and FSH (50 ng/ml D0-D6; 100 ng/ml D7-D12; 200 ng/ml D13-D18) - Sequential FSH - ↑ follicle growth; Act A and FSH alone ↑ follicle survival. |
| Araújo et al., 2014a - α-MEM (3 mg/mL BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 6.7 ng/ml SEL, 2 mM GLUT, 2 mM HYP, 50 µg/ml AA, 100 ng/ml FSH) | Secondary - Isolated 2D/3D (alginate - 32 days) | 100 ng/ml VEGF, 50 ng/ml GH, 50 ng/ml IGF-I - System 2D X 3D - VEGF + 2D: ↑ diameter, antrum formation and growth rate, and maintained morphology |
| Jorssen et al., 2014 - McCoy´s 5a (10mM Hepes, 3mM GLUT; 0.1% BSA, 5 µg/ml TRAN, 5 ng/ml SEL, 5 µg /ml INS, 50 µg/ml AA) | Preantral - In situ (6 days) | 50 µg/ml neutral red (NR) in the presence of: 5% O2 vs. 20% O2 - Follicle dynamics are not influenced by O2 tension |
| Castro et al., 2014 - Base media (3mM GLUT; 2mM HYP, 0.1% BSA, 2.5 µg/ml TRAN, 4 ng/ml SEL, 10 ng/ml INS, 50 µg/ml AA) | Preantral - In situ (5 days) | Different base media: McCoy, α-MEM, and TCM199 (Fresh vs. Vitrified) - TCM199 maintained morphology (Fresh tissue); McCoy ↑ growth and maintained viability (Vitrified tissue) |
| Araújo et al., 2015 - α-MEM or TCM199 (3 mg/mL BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYP, 50 µg/ml AA) | Secondary - Isolated 2D (32 days) | α-MEM vs. TCM199 (60µl-Replacement - C vs. 5 µl-addition - S) - ↑diameter and antrum formation in TCM199-C vs α-MEM-C. |
| Rossetto et al., 2016 - TCM-199-HEPES (3mM GLUT, 0.1% BSA, 2.5 µg/ml TRAN, 4 ng/ml SEL, 10 ng/ml INS, 50 µg/ml AA, 100 ng/ml Act A, 100 ng/ml FSH | Secondary - Isolated 2D and 3D (Collagen - 18 days) | 5, 10 ng/ml or µg/ml INS (2D) and 10 ng/ml INS+ FSH fixed (100 ng/ml) vs. sequential (1 ng/mlD0-D6; 10 ng/mlD6-D12; 100 ng/mlD12-D18) -10 ng/ml INS alone or with fixed FSH ↑ diameter and daily growth. |
| Jimenez et al., 2016 - Base media (20 mM Hepes, 3mM GLUT, 0.1% BSA, 2.5 µg/ml TRAN, 4 ng/ml SEL, 10 ng/ml INS, 50 µg/ml AA | Preantral - In situ (7 days) | Different base media: αMEM, TCM199 or McCoy - αMEM showed better results regarding follicle viability and growth |
| Passos et al., 2016 - α-MEM (2 mM GLUTA, 2 mM HYPO, 3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRANS, 5 ng/ml SEL) | Preantral- In situ (6 days) | 1, 10, 50 and 100 ng/ml IL-1β - 10 ng/ml IL-1β - ↑developing follicles and maintained normal morphology |
| Silva et al., 2017a - α-MEM( 2 mM GLUTA, 2 mM HYPO, 1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRANS, 5 ng/ml SEL) | All follicular categories - In situ (6 days) | 1, 10, 100 or 200 ng/ml TNF-α and 1, 10, 100 or 200 ng/ml dexamethasone - TNF-α ↑ cell apoptosis and 10 ng/ml dexamethasone conserved follicle ultrastructure |
| Zoheir et al., 2017 - TCM-199 (10% Newborn Calf Serum, 0.23 mM PYRU, 1% ITS, 2.2 g/l sodium bicarbonate, 100 ng/ml FSH, 100 ng/ml EGF | Secondary - Isolated 2D (15 days) | 100, 250 or 500 µM ALA - ALA ↑follicle growth and maintained viability |
*All the results are compared to control group. Abbreviations: 2D, two-dimensional culture system; 3D, three-dimensional culture system; BSA, bovine serum albumin; GLUT, glutamine; INS, Insulin; TRAN, Transferrin; SEL, Selenium AA, acid ascorbic; Hypo, Hypoxanthine; PYRU, Pyruvate; FBS, fetal bovine serum; ITS, commercial insulin, transferrin, selenium; FSH, follicle-stimulating hormone; Act A, activin A; GDF-9, growth and differentiation factor 9; bFGF, basic fibroblast growth factor; BMP-15, bone morphogenetic protein 15; LH, luteinizing hormone; EGF, epidermal growth factor; VEGF, vascular endothelial growth factor; IGF-I, insulin-like growth factor; GH, growth hormone; IL-1β, interleukin-1 β; TNF-α, tumor necrosis factor-alpha; ALA, alpha lipoic acid.