Limits...
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.

Show MeSH

Related in: MedlinePlus

Expression of PCNA, phos-AKT and phos-SMAD1/5 in human Sertoli cells after culture for 2 months. Western blots displayed the expression of PCNA, phos-AKT, and phos-SMAD1/5 in human Sertoli cells at P1, P5, P10. ACTB served as the control of loading proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig7: Expression of PCNA, phos-AKT and phos-SMAD1/5 in human Sertoli cells after culture for 2 months. Western blots displayed the expression of PCNA, phos-AKT, and phos-SMAD1/5 in human Sertoli cells at P1, P5, P10. ACTB served as the control of loading proteins.

Mentions: We also compared the proliferation potential and signaling pathways of human Sertoli cells after culture for 1, 5, and 10 passages. Western blots showed that there was no difference in the expression of PCNA in human Sertoli cells at P1, P5, and P10 (Figure 7), and phos-AKT and phos-SMAD1/5 were expressed at similar levels in human Sertoli cells at P1, P5, and P10 (Figure 7). These data suggest that the activation of AKT and SMAD1/5 signaling pathways was maintained in human Sertoli cells during the long-period culture.Figure 7


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)

Expression of PCNA, phos-AKT and phos-SMAD1/5 in human Sertoli cells after culture for 2 months. Western blots displayed the expression of PCNA, phos-AKT, and phos-SMAD1/5 in human Sertoli cells at P1, P5, P10. ACTB served as the control of loading proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig7: Expression of PCNA, phos-AKT and phos-SMAD1/5 in human Sertoli cells after culture for 2 months. Western blots displayed the expression of PCNA, phos-AKT, and phos-SMAD1/5 in human Sertoli cells at P1, P5, P10. ACTB served as the control of loading proteins.
Mentions: We also compared the proliferation potential and signaling pathways of human Sertoli cells after culture for 1, 5, and 10 passages. Western blots showed that there was no difference in the expression of PCNA in human Sertoli cells at P1, P5, and P10 (Figure 7), and phos-AKT and phos-SMAD1/5 were expressed at similar levels in human Sertoli cells at P1, P5, and P10 (Figure 7). These data suggest that the activation of AKT and SMAD1/5 signaling pathways was maintained in human Sertoli cells during the long-period culture.Figure 7

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