Optimal culture conditions are critical for efficient expansion of human testicular somatic and germ cells in vitro

Gat, Itai; Maghen, Leila; Filice, Melissa; Wyse, Brandon A.; Zohni, Khaled; Jarvi, Keith; Lo, Kirk; Fisher, Andrée Gauthier; Librach, Clifford L.

doi:10.1016/j.fertnstert.2016.12.028

Cited by 26 publications

(23 citation statements)

References 31 publications

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“…Kim et al () recognised that STEMPRO medium produced a greater proliferation of SSCs contrasted to DMEM medium in bovine, which would partially agree with our results. Although the comparison of different culture media for the proliferation of SSC has been studied in other species like human (Gat et al, ), this is the first work that analysed the proliferation of alpaca SSCs in different culture media. Medrano et al () observed that STEMPRO promotes proliferation of cryopreserved human SSC, while DMEM did not.…”

Section: Discussionmentioning

confidence: 99%

“…The presence of sodium pyruvate, glutamine, sodium selenite and nonessential amino acids has been observed in culture media in different species, for example, in mice (Guan et al, ; Sato et al, ), porcine (Goel et al, ; Lee et al, ), bovine (Kim et al, ), goat (Pramod & Mitra, ) and human (Gat et al, ), allowing the maintenance of the cells. Some authors have also used BSA and FBS as a source of proteins, amino acids and growth factors that induce cell proliferation (Nayak et al, ).…”

Section: Discussionmentioning

confidence: 99%

“…Spermatogonial stem cell culture allows the maintenance of the lineage, induces cell proliferation and also is useful for the study of the molecular mechanisms involved in the gamete production, improving the knowledge on male fertility in different species. Culture media as DMEM and STEMPRO were used in human (Gat et al ., ), DMEM in mouse (Hamidabadi & Bojnordi, ) and DMEM and KSOM in bovine (Izadyar et al, ), DMEM with high glucose that induces optimal in vitro culture conditions to SSC expansion in domestic cat (Bedford‐Guaus et al, ). Growth factors such as GDNF, FGF and EGF have used in the SSC culture media, which can be added to the medium via foetal bovine serum, which is a component widely used in stem cell culture systems (Bedford‐Guaus et al, ; Kubota, Avarbock, & Brinster, ).…”

Section: Introductionmentioning

confidence: 99%

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In vitro culture of spermatogonial stem cells isolated from adult alpaca (Vicugna pacos) testes analysed withDolichos biflorusby flow cytometry

et al. 2019

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Spermatogonial stem cell (SSC) is known for its self‐renewal capacity. We have studied the in vitro proliferation of isolated SSC from adult alpaca (Vicugna pacos) testes. A total of 107 samples were evaluated of which 31 were evaluated at baseline, 36 were cultivated in DMEM and 40 in STEMPRO media. Half of the cultivated samples was analysed after 14 days, and the rest after 21 days. Round cell subpopulations were identified with FITC‐DBA by flow cytometry: strongly positive DBA (sDBA+) as SSC, weakly positive DBA (wDBA+) as in early differentiation and negative DBA as differentiated. At the beginning, 4.16% of the cells were SSC, 37.61% wDBA+ while 54.12% were DBA−. After 14 days, 42.28% of SSC, 44.68% wDBA+ and 11.07% DBA− were found in DMEM while 47.09% of SSC, 32.57% wDBA+ and 18.48% DBA− in STEMPRO. After 21 days 38.66% were SSC, 52.78% wDBA and 7.65% DBA− in DMEM and on STEMPRO media 47.92% SSC, 44.20% wDBA+, 4.93% DBA−. There is a significant difference between the number of initial and SSC cultivated, as well as between DBA− (p < 0.05), while there is no significant difference between the wDBA+ (p > 0.05). Our results suggest that both culture media are appropriate for the in vitro proliferation of alpacas SSC.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In vitro culture of spermatogonial stem cells isolated from adult alpaca (Vicugna pacos) testes analysed withDolichos biflorusby flow cytometry

et al. 2019

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“…The requirements for optimal SSC culture conditions have been suggested to differ for rodents and human SSCs, and this requires further studies [Medrano et al 2016]. Nonetheless, the initial somatic to germ cell ratio established during primary SSC culture has been shown to impact the efficiency of early human SSC culture [Gat et al 2017].…”

Section: Discussionmentioning

confidence: 99%

Initial germ cell to somatic cell ratio impacts the efficiency of SSC expansion in vitro

Gat

Maghen

Filice

et al. 2017

Systems Biology in Reproductive Medicine

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“…52,53 The influence of the medium was also pinpointed when Gat et al observed more germ-cell aggregate formation when using DMEM/ F12 instead of StemPro-34. 54 Others also examined the efficiency of differential plating to select germ cells from TCSs, but did not find a difference in germ-cell numbers recovered from whole TCSs and differentially plated cells after 14 days of culture. 55 To improve SSC propagation, cell sorting prior to culture was further applied.…”

Section: Ssc Propagationmentioning

confidence: 99%

<p>Role of stem cells in fertility preservation: current insights</p>

Vermeulen¹,

Giudice²,

Vento³

et al. 2019

SCCAA

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While improvements made in the field of cancer therapy allow high survival rates, gonadotoxicity of chemo- and radiotherapy can lead to infertility in male and female pre- and postpubertal patients. Clinical options to preserve fertility before starting gonadotoxic therapies by cryopreserving sperm or oocytes for future use with assisted reproductive technology (ART) are now applied worldwide. Cryopreservation of pre- and postpubertal ovarian tissue containing primordial follicles, though still considered experimental, has already led to the birth of healthy babies after autotransplantation and is performed in an increasing number of centers. For prepubertal boys who do not produce gametes ready for fertilization, cryopreservation of immature testicular tissue (ITT) containing spermatogonial stem cells may be proposed as an experimental strategy with the aim of restoring fertility. Based on achievements in nonhuman primates, autotransplantation of ITT or testicular cell suspensions appears promising to restore fertility of young cancer survivors. So far, whether in two- or three-dimensional culture systems, in vitro maturation of immature male and female gonadal cells or tissue has not demonstrated a capacity to produce safe gametes for ART. Recently, primordial germ cells have been generated from embryonic and induced pluripotent stem cells, but further investigations regarding efficiency and safety are needed. Transplantation of mesenchymal stem cells to improve the vascularization of gonadal tissue grafts, increase the colonization of transplanted cells, and restore the damaged somatic compartment could overcome the current limitations encountered with transplantation.

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Optimal culture conditions are critical for efficient expansion of human testicular somatic and germ cells in vitro

Cited by 26 publications

References 31 publications

In vitro culture of spermatogonial stem cells isolated from adult alpaca (Vicugna pacos) testes analysed withDolichos biflorusby flow cytometry

In vitro culture of spermatogonial stem cells isolated from adult alpaca (Vicugna pacos) testes analysed withDolichos biflorusby flow cytometry

Initial germ cell to somatic cell ratio impacts the efficiency of SSC expansion in vitro

<p>Role of stem cells in fertility preservation: current insights</p>

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