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Table 2 Summarization of all used protocols for isolation of germline stem cells from different species ovary (NS: Not Stated)

From: Used protocols for isolation and propagation of ovarian stem cells, different cells with different traits

Method Author year & reference Species (s) Age Marker OSC traits OLC size Culture duration Putative OSC’s origin Colony shape and formation duration Superiority and limitation Growth factors used
FACS White 2012 [6, 19, 20] Human
Mouse
(C57BL/6)
6–8 weeks mice
20s, 30s, 40s, 50s women
COOH–DDX4 5–8 μm primitive germ cells genetic signature 35–50 μm 18 months Bone Marrow and peripheral blood 10–12 weeks in mouse
4–8 weeks in human
Viable and purified population N-2 supplement,
LIF
EGF
bFGF
GDNF
Virant klun 2008 [29, 30] Human Reproductive ages and menopause SSEA-4 2–4 μm and bigger cells 8 μm small, round and yellow-color 60 μm 21–23 days an integral part of the ovarian surface epithelium approximately 3 months Homogenous population Without growth factors
Dunlop 2014. [79] Bovine Human NS DDX4 NS NS several month NS NS NS NS
OSE Scraping Virant klun 2008 [23, 24] Human Reproductive ages and menopause OSCs separated by density gradient centrifugation Small round cells with a bubble-like structure 95 μm 20 days PGCs and VSELs NS Simple and easy Without growth factors
Bukovsky 2004 [21, 22] Human 27–38 years women Whole ovarian cells were cultured 10 μm nuclear MAPK & PS1 immunoexpression 180 μm 5–6 days mesenchymal Somatic cells in the tunica albuginea NS Simple and easy Without growth factors
Bhartiya 2011 [33] Human
Monkey
Sheep
Rabbit
Menopausal women with a mean age range of 46 years Whole ovarian cells were cultured Smaller PSCs: 1–3 μm
Larger PSCs: 4–7 μm
Dark, bubbly, shiny appearance
Large, darkly stained nuclei with a thin rim of cytoplasm
130 μm 3 weeks VSELs origin Day 10 Flat with a well-defined margin ES cell-like colonies
3-dimensional dense floating embryoid body-like structures
Simple and easy Without growth factors
MACS Johnson 2005 [21] Mouse 6 and 9 weeks SSEA-1 Isolated cells were used directly for RT-PCR Bone Marrow and peripheral blood NS NS NS
Zou 2009 [18] Mouse (C57BL/6) Adult and 5 day old female MVH Large round or ovoid cells with little cytoplasm and spherical nuclei with slight staining, a large ratio of nuclear plasma and nuclear diameter of 12–20 μm NS 15 months and more for neonatal FGSCs
6 months for adults FGSCs
NS After 7–8 passages (forming cluster) Poor describing details
Relatively low efficiency
LIF
Transferrin
Insulin
Putrescine
EGF
GDNF
bFGF
Zou 2011 [43] CD-1 mice Fragilis 5-day-old female NS NS NS NS NS Improved Efficiency LIF
Transferrin
Insulin
Putrescine
EGF
GDNF
bFGF
Zhou 2013 [38, 39] Rat 5-day-old Fragilis Round with a high nuclear to cytoplasm ratio 55–60 um More than one year NS In first 3 weeks: spherical or grape-like clusters consisting of 4–8 cells Better efficiency of gene transfer for producing transgenic rat LIF
EGF
GDNF
bFGF
size and morphology similar to mouse FGSCs
Bui 2014 [69] Pig Prepubertal gilts SSEA-4 5–7 μm Completely round nuclei that took up almost the entire volume of the cell >100 μm 6 months VSELs and epiblast-derived PGCs Spherical colonies comprising compact clusters of small round after 1 day
Dark and shiny colonies
Better culture condition for establishment and long-term maintenance of PSCs B27
SCF
Using ΔPE-Oct4-Gfp transgenic mice Pacchiarotti 2010 [48] Mouse 2 and 5 days old or one adult mouse GFP which is express under Oct-4 promoter 10–15 μm Located in OSE positive for VASA, c-Kit, SSEA-10.05 % GFP positive in adults and 1–2 % in neonata Up to 60 μm More than one year They are reserved pool in the quiescent state One week Is not optimal for several reasons [20] Insulin
Transferrin
Selenium
Fibronectin
Morphology based selection methods Parvari 2015 [50] Mouse 5–7 day old OSCs separated by colony selection 2–4 μm As same as VSELs Slightly bigger than OSCs 11 days VSELs origin 4 days after pre-plating Small colonies were highly compact without a clear border Easy Affordable LIF