Table 1.
Culture conditions for spermatogonial stem cells in human, monkey, mouse, cattle, goat, sheep, and pig
| Subject, reference | Donor age | Isolation method | Enrichment method | Medium | Serum | Growth factors | Feeder cells | Identification method | Duration of culture (day) |
|---|---|---|---|---|---|---|---|---|---|
| Human, Sadri-Ardekani et al.108 | NA | Two-step enzymatic digestion | Differential plating | StemPro-34 serum-free medium | 10% FCS | 20 ng ml−1 EGF, 10 ng ml−1 GDNF, 10 ng ml−1 LIF | NA | Immunostaining | Approximately 196 |
| Human, Guo et al.109 | 22–35 years | Two-step enzymatic digestion | Differential plating + MACS for GPR125 | StemPro-34 serum-free medium | 2.5 mg ml−1 lipid-rich BSA, 1% FBS | 50 ng ml−1 GDNF, 20 ng ml−1 EGF, 10 ng ml−1 bFGF, 10 ng ml−1 LIF | NA | Immunostaining | Approximately 60 |
| Monkey, Langenstroth et al.110 | 3.7–4.6 years | Two-step enzymatic digestion | Differential plating | α-MEM | 10% FBS | NA | Somatic cells | Immunostaining | Approximately 11 |
| Mouse, Kubota et al.28 | 4.5–7.5 days | Two-step enzymatic digestion | MACS for Thy1 | α-MEM | 0.2% BSA | 40 ng ml−1 GDNF, 300 ng ml−1 GFRα1, 1 ng ml−1 bFGF | STO | Immunostaining | Approximately 180 |
| Mouse, Kanatsu-Shinohara et al.111 | 8 days | Two-step enzymatic digestion | MACA for CD9 | StemPro-34 serum-free medium | 3 mg ml−1 of lipid-rich BSA, 1:100 lipid mixture 1, 1:1000 lipoprotein-cholesterol concentrate | 20 ng ml−1 EGF, 10 ng ml−1 FGF2, 15 ng ml−1 GDNF | NA | Flow cytometry | Approximately 178 |
| Cattle, Oatley et al.93 | 4–5 months | Two-step enzymatic digestion | Percoll gradient + differential plating | α-MEM | 0.5% BSA | 20 ng ml−1 GDNF, 2 ng ml−1 FGF2, 100 ng ml−1 LIF | Bovine fetal fibroblasts | Immunostaining | Approximately 60 |
| Cattle, Herrid et al.30 | 5–7 months | Two-step enzymatic digestion | Differential plating | DMEM | 5% FBS | NA | NA | Immunostaining | NA |
| Goat, Ren et al.112 | 2–2.5 months | Two-step enzymatic digestion | Differential plating | DMEM | 5% FBS | 25 ng ml−1 EGF, 20 ng ml−1 PDGF-BB, 5 ng ml−1 bFGF | Sertoli cells | Immunostaining | Approximately 28 |
| Goat, Pramod and Mitra50 | 3–4 months | Two-step enzymatic digestion | Percoll gradient | DMEM | 10% FBS | NA | Sertoli cells | Immunostaining | Approximately 45 |
| Sheep, Binsila et al.37 | Prepubertal | Two-step enzymatic digestion | Differential plating | DMEM | 5% FBS | NA | NA | Immunostaining | NA |
| Sheep, Binsila et al.113 | Prepubertal | Two-step enzymatic digestion | Ficoll density gradient + differential plating | StemPro-34 serum-free medium | 5% FBS | 40 ng ml−1 GDNF, 20 ng ml−1 EGF, 100 ng ml−1 IGF1 | NA | Immunostaining | Approximately 36 |
| Pig, Goel et al.114 | 2–4 days | Two-step enzymatic digestion | Percoll gradient + differential plating | DMEM | 10% FBS | 10 µg ml−1 insulin | NA | Immunostaining | Approximately 7 |
| Pig, Zheng et al.115 | 7 days | Two-step enzymatic digestion | FACS for PLD6 | DMEM | 5% FBS + 5% KSR | 20 ng ml−1 GDNF, 40 ng ml−1 GFRα1, 10 ng ml−1 bFGF | NA | Immunostaining | Approximately 210 |
bFGF: basic fibroblast growth factor; BSA: bovine serum albumin; DMEM: Dulbecco’s modified Eagle’s medium; EGF: epidermal growth factor; FBS: fetal bovine serum; FCS: fetal calf serum; FGF2: fibroblast growth factor 2; GDNF: glial cell line-derived neurotrophic factor; GFRα1: glial cell line-derived neurotrophic factor receptor alpha 1; IGF1: insulin-like growth factor 1; KSR: knockout serum replacement; LIF: leukemia inhibitory factor; MACS: magnetic-activated cell sorting; FACS: fluorescence-activated cell sorting; MEM: minimum essential medium; NA: not available; PDGF-BB: platelet-derived growth factor-BB; STO: SIM mouse embryonic fibroblasts; GPR: G-protein receptor; THY1: Thy1 cell surface antigen; CD9: CD9 molecule; PLD6: phospholipase D family, member 6