Table 3. Chronological advances in in vitro culture of caprine preantral follicles*.
| References/Base medium | Follicle Category/Culture system | Tested compounds/Main findings |
| Saraiva et al., 2010 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH) | Secondary - Isolated 2D (18 days) | 50 or 100 ng/ml LH and 50, or 100 ng/ml EGF alone or associated - LH+EGF ↑ follicle growth and oocyte meiotic resumption. 2 embryos (8- and 16-cell) in 100 ng/ml LH+EGF |
| Saraiva et al., 2011 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Secondary - Isolated 2D (18 days) | Fixed (100, or 1000 ng/ml), Sequential FSH - Sequential FSH ↓extrusion, ↑ antrum formation and oocyte meiotic resumption. |
| Magalhães et al., 2011 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH) | Secondary - Isolated 2D (18 days) | 10 or 50 ng/ml GH - 50 ng/ml GH ↑ antrum formation and oocyte meiotic resumption. 1 morula from 50 ng/ml GH |
| Araújo et al., 2011 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH) | Secondary - Isolated 2D (18 days) | 10 or 100 ng/ml VEGF - 100 ng/ml VEGF ↑ oocyte meiotic resumption |
| Chaves et al., 2012 - α-MEM (3 mg/ml BSA, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Secondary - Isolated 2D (18 days) | 5 or 10 ng/ml, or 10 µg/ml insulin alone or in association to sequential - 10 ng/ml insulin with sequential FSH ↑ oocyte meiotic resumption |
| Lima et al., 2012 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Primordial and primary - In situ (16 days) | 50 ng/ml KL and 50 ng/ml FSH alone or in different combinations - KL (D0-D8)/FSH(D8-D16) ↑activation and growth |
| Magalhães-Padilha et al., 2012 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Primordial and primary - In situ (16 days) | 50 ng/ml IGF-I and 50 ng/ml FSH alone, or in different combinations - IGF-I+FSH ↑activation and growth, number of secondary follicles |
| Magalhães-Padilha et al., 2012a - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Primordial and primary - In situ (16 days) | 10 ng/ml GH and 50 ng/ml FSH alone, or in different combinations - FSH (D0-D8)/GH (D8-D16) ↑ follicle morphology, viability, activation, growth and secondary follicles |
| Magalhães-Padilha et al., 2012b - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH) | Secondary - Isolated 2D (18 days) | 50 or 100 ng/ml IGF-I - IGF-I ↑ oocyte meiotic resumption. 50 ng/ml IGF-I ↑ antrum formation |
| Cunha et al., 2013 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, 100 ng/ml FSH) | Secondary - Isolated 2D (18 days) | 1, 10, 50, 100 or 200 µg/ml PHA-10 µg/ml PHA maintained follicle ultrastructure and ↑antrum formation |
| Duarte et al., 2013 - α-MEM (3 mg/ml BSA, 10 ng/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Secondary - Isolated 2D (18 days) | 20 or 50 ng/ml IGF-II, or sequential FSH - 20 ng/ml IGF-II alone or in associated to sequential FSH ↑ oocyte meiotic resumption † |
| Faustino et al., 2013 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Primordial and primary - In situ (7 days) | 1 ng/ml KGF-1 and 50 ng/ml KL alone or in combination - 50 ng/ml KL alone ↑ activation and growth |
| Rocha et al., 2013 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 0.23mM PYR) | Primordial and primary - In situ (7 days) | 100, 250, 500, or 1000 pM melatonin, and 50 ng/ml FSH, alone or in combination - 1000 pM melatonin+50 ng/ml FSH ↑ follicular and oocyte diameters |
| Silva et al., 2013 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH) | Secondary - Isolated 2D (18 days) | 50 or 100 ng/ml EGF - EGF ↑ follicle daily growth rate. 50 ng/ml EGF ↑ oocyte meiotic resumption |
| Amin et al., 2013 - Bicarbonate-buffered TCM199 | Secondary - Isolated 2D (6 days) | Different concentrations and association of T4, FSH, GH, EGF, and IGF-I - All substances alone or in combination ↑ follicle growth and antrum formation. The association T4+FSH+GH+EGF ↑ extrusion. |
| References/Base medium | Follicle Category/Culture system | Tested compounds/Main findings |
| Pessoa et al., 2014 - α-MEM (3 mg/ml BSA, 10 ng/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH and 50 ng/ml GH) | Secondary - Isolated 2D (Different days) and 3D (alginate - 36 days) | Different culture periods: 18, 24, 30, 36, or 42 days (2D vs. 3D) - Follicle diameter and oocyte meiotic resumption ↑ until day 36. 3D system did not affect oocyte meiotic resumption. |
| Portela et al., 2014 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 0.23mM PYR) | Primordial and primary - In situ (6 days) | 5, 10, 20 or 10 µg/ml Con A. 10 µg/ml Con A and 50 ng/ml FSH, alone or in combination - 10 µg/ml Con A and 50 ng/ml FSH alone ↑activation. |
| 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 HYPO, 50 µg/ml AA, Sequential FSH, 100 ng/ml VEGF, 1 mg/ml FET ) | Secondary - Isolated 2D and Isolated 3D (0.5% alginate - 18 days) | Follicles from Adult Vs. Prepubertal ovaries (2D vs. 3D) - 3D system ↑ survival and ↓ oocyte extrusion. Greater follicle and oocyte and meiotic resumption diameter in 2D from adult ovaries. Four 2-cell embryos in 2D and one 8 cell-embryo in 3D. |
| Rocha 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 HYPO, 50 µg/ml AA, 100 ng/ml VEGF, 1 mg/ml FET) | Secondary - Isolated 3D (0.5% alginate - 18 days) | 10 mIU/mlvhuman FSH Vs. Sequential FSH - 10 mIU/ml human FSH ↑oocyte meiotic resumption. |
| Brito 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 HYPO, 50 µg/ml AA, Sequential FSH ) | Secondary - Isolated 3D (alginate - 18 days) | 0.25%, 0.5%, or 1% alginate - 0.5% alginate maintained better follicle integrity. 0.25% alginate ↑ follicle diameter, daily growth and oocyte meiotic resumption. |
| Almeida et al., 2015 - α-MEM (1.25 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 AA) | Primordial and primary - In situ (16 days) | 50 ng/ml FGF-10 and 50 ng/ml FSH alone, in combination or sequentially - FSH(D0-D8)/FGF-10 (D8-D16) showed ↑ percentages of normal and growing follicles |
| Lopes et al., 2015 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) | Primordial and primary - In situ (6 days) | 10 µg/ml PHA and 100 µg/ml EGF alone or in combination (Healthy goats Vs. CAEV) - EGF alone ↑ growth in both healthy and CAEV infected goats |
| Gouveia et al., 2016 - α-MEM or different concentrations of Amb extracts | Secondary - Isolated 2D (12 days) | α-MEM Vs. Amb extracts (0.1, 0.2 or 0.4 mg/ml). α-MEM, 0.2 mg/ml Amb (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA) or sequential FSH - 100ng/mlD0-D6, 500 ng/mlD6-D12,) - α-MEM and 0.2 mg/ml Amb similar morphology, antrum formation and follicle diameter |
| Ferreira et al., 2016 - α-MEM (3 mg/ml BSA, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA and 50 ng/ml GH) | Secondary - Isolated 2D (18 days) | 10 ng/ml or 10 µg/ml insulin alone or associated to either fixed 100 ng/ml FSH or sequential FSH - 10 ng/ml insulin ↓extrusion. 10 µg/ml insulin ↑ follicle growth. 100 ng/ml FSH ↑oocyte meiotic resumption. 10 µg/ml insulin +100 ng/ml FSH ↑ mean oocyte diameter |
| Brito et al., 2016 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA, Sequential FSH) | Secondary - Isolated 3D (alginate - 18 days) | Number of follicles per 0.25% alginate bead and beads per well. 5 follicles/bead in alginate, fibrin-alginate, or hyaluronate) - 5 follicles/bead ↑diameter. Alginate ↑follicle daily growth. Fibrin-alginate ↑ oocyte meiotic resumption. |
| Silva et al., 2017b - α-MEM (3 mg/ml BSA, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO, 50 µg/ml AA and 100 ng/ml VEGF) | Secondary - Isolated 2D (18 days) | 10 ng/ml or 10 µg/ml insulin alone or associated to either fixed 100 ng/ml FSH or sequential - 10 µg/ml insulin ↑ follicle growth. 10 µg/ml insulin + fixed FSH ↑ oocyte meiotic resumption. |
| Cadenas et al., 2017 - α-MEM (3 mg/ml BSA, 10 ng/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO and 50 µg/ml AA) | Secondary and tertiary - Isolated 2D (24 days - PFs and 18 days EAFs) | 50 ng/ml GH or 100 ng/ml VEGF alone, in combination or sequentially (Secondary PF vs. Tertiary EAF) - 50 ng/ml GH ↑ oocyte growth and meiotic resumption only in EAFs. |
| Ferreira et al., 2018 - α-MEM (3 mg/ml BSA, 10 µg/ml INS, 5.5 µg/ml TRAN, 5 ng/ml SEL, 2 mM GLUT, 2 mM HYPO and 50 µg/ml AA) | Secondary - Isolated 2D (18 days) | 10, 50 or 100 mIU/ml human FSH (PFs vs. EAFS) - No positive effect of human FSH on PFs. On EAFs:50 mIU/ml human FSH ↑ follicle and oocyte growth |
*All the results are compared to control group. Abbreviations: 2D, two-dimensional culture system; 3D, three-dimensional culture system; PFs, preantral follicles; EAFs, early antral follicles; BSA, bovine serum albumin; GLUT, glutamine; INS, Insulin; TRAN, Transferrin; SEL, Selenium AA, acid ascorbic; Hypo, Hypoxanthine; PYRU, Pyruvate; FET, fetuin; LH, luteinizing hormone; EGF, epidermal growth factor; FSH, follicle-stimulating hormone; Sequential FSH - 100 ng/ml in D0-D6, 500 ng/ml in D6-D12,1000 ng/ml in D12-D18; GH, growth hormone; VEGF, vascular endothelial growth factor; KL, Kit ligand; IGF-I, insulin-like growth factor I; IGF-II, insulin-like growth factor II; PHA, phytohemagglutinin; KGF-1, keratinocyte growth factor 1; T4, thyroxine; Con A, concavalin A; FGF-10, fibroblastic growth factor 10; CAEV, caprine arthritis encephalitis lentivirus; Amb, Amburana cearensis extract.