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Article ; Online: Effects of Growth Factors on In Vitro Culture of Neonatal Piglet Testicular Tissue Fragments.

Ibtisham, Fahar / Cham, Tat-Chuan / Fayaz, Mohammad Amin / Honaramooz, Ali

2023 Volume 12, Issue 18

Abstract: In vitro spermatogenesis (IVS) has important applications including fertility preservation of prepubertal cancer patients; however, thus far, IVS has only been achieved using mouse models. To study the effects of growth factors on the maintenance of ... ...

Abstract	In vitro spermatogenesis (IVS) has important applications including fertility preservation of prepubertal cancer patients; however, thus far, IVS has only been achieved using mouse models. To study the effects of growth factors on the maintenance of testicular tissue integrity, germ cell numbers, and potential induction of IVS using a porcine model, we cultured small testicular fragments (~2 mg) from 1-wk-old piglets under six different media conditions (DMEM + 10%KSR alone or supplemented with GDNF, bFGF, SCF, EGF, or a combination of all) for 8 weeks. Overall, tissues supplemented with GDNF and bFGF had the greatest seminiferous tubule integrity and least number of apoptotic cells. GDNF-supplemented tissues had the greatest number of gonocytes per tubule, followed by bFGF-supplemented tissues. There was evidence of gradual Sertoli cell maturation in all groups. Moreover, histological examination and the expression of c-KIT (a marker of differentiating spermatogonia and spermatocytes) and STRA8 (a marker of the pre/meiotic stage germ cells) confirmed the induction of IVS in all groups. However, GDNF- and bFGF-supplemented tissue cultures had greater numbers of seminiferous tubules with spermatocytes compared to other groups. In conclusion, overall, GDNF and bFGF supplementation better maintained the tissue integrity and gonocyte numbers and induced IVS in cultured testicular tissues.
Language	English
Publishing date	2023-09-08
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12182234
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Right atrioventricular valve dysplasia in a meerkat.

Cham, Tat-Chuan / Desprez, Isabelle / Gupta, Ashish / Aburto, Enrique

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

2022 Volume 34, Issue 6, Page(s) 1032–1034

Abstract: A 7-y-old, captive, intact female meerkat ( ...

Abstract	A 7-y-old, captive, intact female meerkat (
MeSH term(s)	Female ; Animals ; Chordae Tendineae/pathology ; Tricuspid Valve ; Heart Failure/veterinary ; Autopsy/veterinary
Language	English
Publishing date	2022-10-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	287603-6
ISSN	1943-4936 ; 1040-6387
ISSN (online)	1943-4936
ISSN	1040-6387
DOI	10.1177/10406387221128208
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Article ; Online: Long-Term In Vitro Maintenance of Piglet Testicular Tissue

Fahar Ibtisham / Tat-Chuan Cham / Mohammad Amin Fayaz / Ali Honaramooz

Animals, Vol 13, Iss 1, p

Effects of Tissue Fragment Size, Preparation Method, and Serum Source

2022 Volume 128

Abstract: Long-term culture of testicular tissue has important applications, including the preservation of fertility potential of prepubertal boys undergoing gonadotoxic cancer treatment. This study was designed to define optimal conditions for the long-term ... ...

Abstract	Long-term culture of testicular tissue has important applications, including the preservation of fertility potential of prepubertal boys undergoing gonadotoxic cancer treatment. This study was designed to define optimal conditions for the long-term culture of neonatal porcine testicular tissue as an animal model for preadolescent individuals. Testes from 1 wk old donor piglets were used to examine the effects of tissue fragment size (~2, 4, 6, or 8 mg), preparation method (intact, semi-digested, or physically dispersed fragments), and serum source in the media (fetal bovine serum—FBS—or knockout serum replacement—KSR). Testicular fragments were examined weekly for 4 weeks for tissue integrity, seminiferous cord density and morphology, and gonocyte counts. Testicular tissue integrity was dependent on fragment size and preparation method, where the smallest size (2 mg, p < 0.05) and intact preparation method were advantageous ( p < 0.05). Seminiferous cord density decreased over the culture period ( p < 0.05). Although the relative number of gonocytes decreased over time for all sizes and methods ( p < 0.01), smaller intact fragments (2 and 4 mg) had greater numbers of gonocytes ( p < 0.05). Our findings suggest that intact or physically dispersed testicular fragments of the smallest size (2 mg) cultured in KSR-supplemented media could be effectively maintained in vitro for the duration of 4 weeks.
Keywords	in vitro spermatogenesis ; tissue culture ; infertility ; organ culture systems ; porcine testicular tissue ; Veterinary medicine ; SF600-1100 ; Zoology ; QL1-991
Subject code	630
Language	English
Publishing date	2022-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Culture supplementation of bFGF, GDNF, and LIF alters in vitro proliferation, colony formation, and pluripotency of neonatal porcine germ cells.

Fayaz, Mohammad Amin / Ibtisham, Fahar / Cham, Tat-Chuan / Honaramooz, Ali

Cell and tissue research

2022 Volume 388, Issue 1, Page(s) 195–210

Abstract: Gonocytes in the neonatal testis have male germline stem cell properties and as such have important potential applications in fertility preservation and regenerative medicine. Such applications require further studies aimed at increasing gonocyte numbers ...

Abstract	Gonocytes in the neonatal testis have male germline stem cell properties and as such have important potential applications in fertility preservation and regenerative medicine. Such applications require further studies aimed at increasing gonocyte numbers and evaluating their pluripotency in vitro. The objective of the present study was to test the effects of basic fibroblast growth factor (bFGF), glial cell line-derived neurotrophic factor (GDNF), and leukemia inhibitory factor (LIF) on in vitro propagation, colony formation, and expression of pluripotency markers of neonatal porcine gonocytes. Testis cells from 1-week-old piglets were cultured in basic media (DMEM + 15% FBS), supplemented with various concentrations of bFGF, GDNF, and LIF, either individually or in combinations, in a stepwise experimental design. Gonocytes and/or their colonies were evaluated every 7 days and the gonocyte- (DBA) and pluripotency-specific markers (POU5F1, SSEA-1, E-cadherin, and NANOG) assessed on day 28. Greatest gonocyte numbers and largest colonies were found in media supplemented with 10 ng/mL bFGF and 10 ng/mL bFGF + 100 ng/mL GDNF + 1500 U/mL LIF, respectively. The resultant gonocytes and colonies expressed both germ cell- and pluripotency-specific markers. These results shed light on the growth hormone requirements of porcine gonocytes for in vitro proliferation and colony formation.
MeSH term(s)	Animals ; Cell Proliferation ; Cells, Cultured ; Dietary Supplements ; Fibroblast Growth Factor 2/pharmacology ; Germ Cells ; Glial Cell Line-Derived Neurotrophic Factor/pharmacology ; Leukemia Inhibitory Factor/pharmacology ; Male ; Swine
Chemical Substances	Glial Cell Line-Derived Neurotrophic Factor ; Leukemia Inhibitory Factor ; Fibroblast Growth Factor 2 (103107-01-3)
Language	English
Publishing date	2022-02-01
Publishing country	Germany
Document type	Journal Article
ZDB-ID	125067-x
ISSN	1432-0878 ; 0302-766X
ISSN (online)	1432-0878
ISSN	0302-766X
DOI	10.1007/s00441-022-03583-3
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Article: Long-Term In Vitro Maintenance of Piglet Testicular Tissue: Effects of Tissue Fragment Size, Preparation Method, and Serum Source.

Ibtisham, Fahar / Cham, Tat-Chuan / Fayaz, Mohammad Amin / Honaramooz, Ali

Animals : an open access journal from MDPI

2022 Volume 13, Issue 1

Abstract	Long-term culture of testicular tissue has important applications, including the preservation of fertility potential of prepubertal boys undergoing gonadotoxic cancer treatment. This study was designed to define optimal conditions for the long-term culture of neonatal porcine testicular tissue as an animal model for preadolescent individuals. Testes from 1 wk old donor piglets were used to examine the effects of tissue fragment size (~2, 4, 6, or 8 mg), preparation method (intact, semi-digested, or physically dispersed fragments), and serum source in the media (fetal bovine serum—FBS—or knockout serum replacement—KSR). Testicular fragments were examined weekly for 4 weeks for tissue integrity, seminiferous cord density and morphology, and gonocyte counts. Testicular tissue integrity was dependent on fragment size and preparation method, where the smallest size (2 mg, p < 0.05) and intact preparation method were advantageous (p < 0.05). Seminiferous cord density decreased over the culture period (p < 0.05). Although the relative number of gonocytes decreased over time for all sizes and methods (p < 0.01), smaller intact fragments (2 and 4 mg) had greater numbers of gonocytes (p < 0.05). Our findings suggest that intact or physically dispersed testicular fragments of the smallest size (2 mg) cultured in KSR-supplemented media could be effectively maintained in vitro for the duration of 4 weeks.
Language	English
Publishing date	2022-12-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13010128
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Article ; Online: Current progress, challenges, and future prospects of testis organoids†.

Cham, Tat-Chuan / Chen, Xiongbiao / Honaramooz, Ali

Biology of reproduction

2021 Volume 104, Issue 5, Page(s) 942–961

Abstract: Spermatogenic failure is believed to be a major cause of male infertility. The establishment of a testis organoid model would facilitate the study of such pathological mechanisms and open the possibility of male fertility preservation. Because of the ... ...

Abstract	Spermatogenic failure is believed to be a major cause of male infertility. The establishment of a testis organoid model would facilitate the study of such pathological mechanisms and open the possibility of male fertility preservation. Because of the complex structures and cellular events occurring within the testis, the establishment of a compartmentalized testis organoid with a complete spermatogenic cycle remains a challenge in all species. Since the late 20th century, a great variety of scaffold-based and scaffold-free testis cell culture systems have been established to recapitulate de novo testis organogenesis and in vitro spermatogenesis. The utilization of the hydrogel scaffolds provides a 3D microenvironment for testis cell growth and development, facilitating the reconstruction of de novo testis tissue-like structures and spermatogenic differentiation. Using a combination of different strategies, including the use of various scaffolding biomaterials, the incorporation of the living cells with high self-assembling capacity, and the integration of the advanced fabrication techniques, a scaffold-based testis organoid with a compartmentalized structure that supports in vitro spermatogenesis may be achieved. This article briefly reviews the current progress in the development of scaffold-based testis organoids while focusing on the scaffolding biomaterials (hydrogels), cell sources, and scaffolding approaches. Key challenges in current organoid studies are also discussed along with recommendations for future research.
MeSH term(s)	Animals ; Humans ; Male ; Mammals/physiology ; Organoids/cytology ; Testis/cytology
Language	English
Publishing date	2021-01-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1118-6
ISSN	1529-7268 ; 0006-3363
ISSN (online)	1529-7268
ISSN	0006-3363
DOI	10.1093/biolre/ioab014
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Article ; Online: Generation of a Highly Biomimetic Organoid, Including Vasculature, Resembling the Native Immature Testis Tissue

Tat-Chuan Cham / Fahar Ibtisham / Mohammad Amin Fayaz / Ali Honaramooz

Cells, Vol 10, Iss 1696, p

2021 Volume 1696

Abstract: The creation of a testis organoid (artificial testis tissue) with sufficient resemblance to the complex form and function of the innate testis remains challenging, especially using non-rodent donor cells. Here, we report the generation of an organoid ... ...

Abstract	The creation of a testis organoid (artificial testis tissue) with sufficient resemblance to the complex form and function of the innate testis remains challenging, especially using non-rodent donor cells. Here, we report the generation of an organoid culture system with striking biomimicry of the native immature testis tissue, including vasculature. Using piglet testis cells as starting material, we optimized conditions for the formation of cell spheroids, followed by long-term culture in an air–liquid interface system. Both fresh and frozen-thawed cells were fully capable of self-reassembly into stable testis organoids consisting of tubular and interstitial compartments, with all major cell types and structural details expected in normal testis tissue. Surprisingly, our organoids also developed vascular structures; a phenomenon that has not been reported in any other culture system. In addition, germ cells do not decline over time, and Leydig cells release testosterone, hence providing a robust, tunable system for diverse basic and applied applications.
Keywords	testis organoid ; de novo testis organogenesis ; tubulogenesis ; testis cell self-assembly ; Biology (General) ; QH301-705.5
Subject code	571
Language	English
Publishing date	2021-07-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Right atrioventricular valve dysplasia in a meerkat

Cham, Tat-Chuan / Desprez, Isabelle / Gupta, Ashish / Aburto, Enrique

Journal of Veterinary Diagnostic Investigation. 2022 Nov., v. 34, no. 6 p.1032-1034

2022

Abstract: A 7-y-old, captive, intact female meerkat (Suricata suricatta) was presented with lethargy, decreased appetite, dyspnea, and distended abdomen. At autopsy, the right atrium was markedly dilated, and the right atrioventricular valve (RAV) was dysplastic ... ...

Abstract	A 7-y-old, captive, intact female meerkat (Suricata suricatta) was presented with lethargy, decreased appetite, dyspnea, and distended abdomen. At autopsy, the right atrium was markedly dilated, and the right atrioventricular valve (RAV) was dysplastic with shortened or absent chordae tendineae and direct attachment of the valve to the papillary muscles, which, in turn, were fused and abnormally positioned. The right ventricle was considered to be hypertrophied. Also present were hydrothorax, ascites, atelectasis, and hepatic congestion. A diagnosis of RAV dysplasia was made. Histologic findings included hypertrophy of cardiomyocytes and marked centrilobular hepatic congestion and hemorrhage, which were consistent with right-sided heart failure.
Keywords	Suricata suricatta ; abdomen ; appetite ; ascites ; cardiomyocytes ; dyspnea ; females ; heart failure ; hemorrhage ; histology ; hypertrophy ; necropsy ; congenital heart defect ; meerkats ; right atrioventricular valve dysplasia
Language	English
Dates of publication	2022-11
Size	p. 1032-1034.
Publishing place	SAGE Publications
Document type	Article ; Online
ZDB-ID	287603-6
ISSN	1943-4936 ; 1040-6387
ISSN (online)	1943-4936
ISSN	1040-6387
DOI	10.1177/10406387221128208
Database	NAL-Catalogue (AGRICOLA)

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Zs.A 3645: Show issues

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Jg. 1995 - 2021: Lesesall (2.OG)
ab Jg. 2022: Lesesaal (EG)

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Article ; Online: Generation of a Highly Biomimetic Organoid, Including Vasculature, Resembling the Native Immature Testis Tissue.

Cham, Tat-Chuan / Ibtisham, Fahar / Fayaz, Mohammad Amin / Honaramooz, Ali

Cells

2021 Volume 10, Issue 7

Abstract	The creation of a testis organoid (artificial testis tissue) with sufficient resemblance to the complex form and function of the innate testis remains challenging, especially using non-rodent donor cells. Here, we report the generation of an organoid culture system with striking biomimicry of the native immature testis tissue, including vasculature. Using piglet testis cells as starting material, we optimized conditions for the formation of cell spheroids, followed by long-term culture in an air-liquid interface system. Both fresh and frozen-thawed cells were fully capable of self-reassembly into stable testis organoids consisting of tubular and interstitial compartments, with all major cell types and structural details expected in normal testis tissue. Surprisingly, our organoids also developed vascular structures; a phenomenon that has not been reported in any other culture system. In addition, germ cells do not decline over time, and Leydig cells release testosterone, hence providing a robust, tunable system for diverse basic and applied applications.
MeSH term(s)	Animals ; Biomimetic Materials/pharmacology ; Cell Count ; Cryopreservation ; Leydig Cells/cytology ; Leydig Cells/drug effects ; Luteinizing Hormone/metabolism ; Male ; Neovascularization, Physiologic/drug effects ; Organ Specificity ; Organoids/cytology ; Organoids/drug effects ; Organoids/physiology ; Organoids/ultrastructure ; Swine ; Testis/blood supply ; Testis/cytology ; Testis/ultrastructure ; Testosterone/metabolism
Chemical Substances	Testosterone (3XMK78S47O) ; Luteinizing Hormone (9002-67-9)
Language	English
Publishing date	2021-07-05
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells10071696
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Long-Term In Vitro Maintenance of Piglet Testicular Tissue: Effects of Tissue Fragment Size, Preparation Method, and Serum Source

Ibtisham, Fahar / Cham, Tat-Chuan / Fayaz, Mohammad Amin / Honaramooz, Ali

Animals. 2022 Dec. 29, v. 13, no. 1

2022

Abstract	Long-term culture of testicular tissue has important applications, including the preservation of fertility potential of prepubertal boys undergoing gonadotoxic cancer treatment. This study was designed to define optimal conditions for the long-term culture of neonatal porcine testicular tissue as an animal model for preadolescent individuals. Testes from 1 wk old donor piglets were used to examine the effects of tissue fragment size (~2, 4, 6, or 8 mg), preparation method (intact, semi-digested, or physically dispersed fragments), and serum source in the media (fetal bovine serum—FBS—or knockout serum replacement—KSR). Testicular fragments were examined weekly for 4 weeks for tissue integrity, seminiferous cord density and morphology, and gonocyte counts. Testicular tissue integrity was dependent on fragment size and preparation method, where the smallest size (2 mg, p < 0.05) and intact preparation method were advantageous (p < 0.05). Seminiferous cord density decreased over the culture period (p < 0.05). Although the relative number of gonocytes decreased over time for all sizes and methods (p < 0.01), smaller intact fragments (2 and 4 mg) had greater numbers of gonocytes (p < 0.05). Our findings suggest that intact or physically dispersed testicular fragments of the smallest size (2 mg) cultured in KSR-supplemented media could be effectively maintained in vitro for the duration of 4 weeks.
Keywords	animal models ; blood serum ; cancer therapy ; cattle ; piglets ; puberty ; testes
Language	English
Dates of publication	2022-1229
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article ; Online
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13010128
Database	NAL-Catalogue (AGRICOLA)

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