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Long-term culture and significant expansion of human Sertoli cells whilst maintaining stable global phenotype and AKT and SMAD1/5 activation.

Guo Y, Hai Y, Yao C, Chen Z, Hou J, Li Z, He Z - Cell Commun. Signal (2015)

Bottom Line: Morphology, phenotypic characteristics, and the signaling pathways of adult human Sertoli cells from different passages were compared.This study demonstrates that adult human Sertoli cells can be cultured for a long period and expanded with remarkable increase of cell numbers whilst maintaining their primary morphology, phenotype and signaling pathways.This study could provide adequate human Sertoli cells for reproductive and regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. kaoyanyong1@126.com.

ABSTRACT

Background: Sertoli cells play key roles in regulating spermatogenesis and testis development by providing structural and nutritional supports. Recent studies demonstrate that Sertoli cells can be converted into functional neural stem cells. Adult Sertoli cells have previously been considered the terminally differentiated cells with a fixed and unmodifiable population after puberty. However, this concept has been challenged. Since the number of adult human Sertoli cells is limited, it is essential to culture these cells for a long period and expand them to obtain sufficient cells for their basic research and clinic applications. Nevertheless, the studies on human Sertoli cells are restricted, because it is difficult to get access to human testis tissues.

Results: Here we isolated adult human Sertoli cells with a high purity and viability from obstructive azoospermia patients with normal spermatogenesis. Adult human Sertoli cells were cultured with DMEM/F12 and fetal bovine serum for 2 months, and they could be expanded with a 59,049-fold increase of cell numbers. Morphology, phenotypic characteristics, and the signaling pathways of adult human Sertoli cells from different passages were compared. Significantly, adult human Sertoli cells assumed similar morphological features, stable global gene expression profiles and numerous proteins, and activation of AKT and SMAD1/5 during long-period culture.

Conclusions: This study demonstrates that adult human Sertoli cells can be cultured for a long period and expanded with remarkable increase of cell numbers whilst maintaining their primary morphology, phenotype and signaling pathways. This study could provide adequate human Sertoli cells for reproductive and regenerative medicine.

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Related in: MedlinePlus

Microarray analysis revealed global gene expression patterns in human Sertoli cells at different passages. (A-C) Scatter plots displayed the large scale gene expression profiles in human Sertoli cells at P5/P1 (A), P10/P1 (B), and P10/P5 (C). Each dot presented one gene or the expressed sequence tags (EST).
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Fig4: Microarray analysis revealed global gene expression patterns in human Sertoli cells at different passages. (A-C) Scatter plots displayed the large scale gene expression profiles in human Sertoli cells at P5/P1 (A), P10/P1 (B), and P10/P5 (C). Each dot presented one gene or the expressed sequence tags (EST).

Mentions: We next compared global gene expression profiling of human Sertoli cells at P1, P5 and P10 using microarray analysis. To this end, total RNA was extracted from human Sertoli cells at different passages, and gel imaging and electropherograms by Agilent bioanalyzer assays displayed that RNA was of great quality (Additional file 3: Figure S3A-E). Microarray analysis revealed that totally 31,743 genes and the expressed sequence tags (EST) were detected in human Sertoli cells (Figure 4A-C). There were 597 (1.88%), 663 (2.09%), and 177 (0.56%) differentially expressed genes (up-regulated or down-regulated with 2.0 folds or more) in Sertoli cells at P5/P1 (Figure 4A, Table 2), P10/P1 (Figure 4B, Table 2), and P10/P5 (Figure 4C, Table 2), respectively. Meanwhile, a number of important genes for Sertoli cells, including WT1, GATA4, GATA1, GDNF, BMP4, KITLG (also called SCF), FGF2, EGF, FSHR, AR and ABP, were stably expressed in human Sertoli cells after culture for P1, P5 and P10, since their expression was changed with less than 2.0 folds (Table 3).Figure 4


Long-term culture and significant expansion of human Sertoli cells whilst maintaining stable global phenotype and AKT and SMAD1/5 activation.

Guo Y, Hai Y, Yao C, Chen Z, Hou J, Li Z, He Z - Cell Commun. Signal (2015)

Microarray analysis revealed global gene expression patterns in human Sertoli cells at different passages. (A-C) Scatter plots displayed the large scale gene expression profiles in human Sertoli cells at P5/P1 (A), P10/P1 (B), and P10/P5 (C). Each dot presented one gene or the expressed sequence tags (EST).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4380114&req=5

Fig4: Microarray analysis revealed global gene expression patterns in human Sertoli cells at different passages. (A-C) Scatter plots displayed the large scale gene expression profiles in human Sertoli cells at P5/P1 (A), P10/P1 (B), and P10/P5 (C). Each dot presented one gene or the expressed sequence tags (EST).
Mentions: We next compared global gene expression profiling of human Sertoli cells at P1, P5 and P10 using microarray analysis. To this end, total RNA was extracted from human Sertoli cells at different passages, and gel imaging and electropherograms by Agilent bioanalyzer assays displayed that RNA was of great quality (Additional file 3: Figure S3A-E). Microarray analysis revealed that totally 31,743 genes and the expressed sequence tags (EST) were detected in human Sertoli cells (Figure 4A-C). There were 597 (1.88%), 663 (2.09%), and 177 (0.56%) differentially expressed genes (up-regulated or down-regulated with 2.0 folds or more) in Sertoli cells at P5/P1 (Figure 4A, Table 2), P10/P1 (Figure 4B, Table 2), and P10/P5 (Figure 4C, Table 2), respectively. Meanwhile, a number of important genes for Sertoli cells, including WT1, GATA4, GATA1, GDNF, BMP4, KITLG (also called SCF), FGF2, EGF, FSHR, AR and ABP, were stably expressed in human Sertoli cells after culture for P1, P5 and P10, since their expression was changed with less than 2.0 folds (Table 3).Figure 4

Bottom Line: Morphology, phenotypic characteristics, and the signaling pathways of adult human Sertoli cells from different passages were compared.This study demonstrates that adult human Sertoli cells can be cultured for a long period and expanded with remarkable increase of cell numbers whilst maintaining their primary morphology, phenotype and signaling pathways.This study could provide adequate human Sertoli cells for reproductive and regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 160 Pujian Road, Shanghai, 200127, China. kaoyanyong1@126.com.

ABSTRACT

Background: Sertoli cells play key roles in regulating spermatogenesis and testis development by providing structural and nutritional supports. Recent studies demonstrate that Sertoli cells can be converted into functional neural stem cells. Adult Sertoli cells have previously been considered the terminally differentiated cells with a fixed and unmodifiable population after puberty. However, this concept has been challenged. Since the number of adult human Sertoli cells is limited, it is essential to culture these cells for a long period and expand them to obtain sufficient cells for their basic research and clinic applications. Nevertheless, the studies on human Sertoli cells are restricted, because it is difficult to get access to human testis tissues.

Results: Here we isolated adult human Sertoli cells with a high purity and viability from obstructive azoospermia patients with normal spermatogenesis. Adult human Sertoli cells were cultured with DMEM/F12 and fetal bovine serum for 2 months, and they could be expanded with a 59,049-fold increase of cell numbers. Morphology, phenotypic characteristics, and the signaling pathways of adult human Sertoli cells from different passages were compared. Significantly, adult human Sertoli cells assumed similar morphological features, stable global gene expression profiles and numerous proteins, and activation of AKT and SMAD1/5 during long-period culture.

Conclusions: This study demonstrates that adult human Sertoli cells can be cultured for a long period and expanded with remarkable increase of cell numbers whilst maintaining their primary morphology, phenotype and signaling pathways. This study could provide adequate human Sertoli cells for reproductive and regenerative medicine.

Show MeSH
Related in: MedlinePlus