Alteration in genes expression patterns during in vitro differentiation of mouse spermatogonial cells into neuroepithelial-like cells

Abstract

Pluripotent stem cells derived from testis is a new, natural, and unlimited source for cell therapy in regenerative medicine and represent a possible alternative to replacing of all cells in the body. Here, we designed a simple co-culture system of spermatogonia cells with Sertoli cells for the generation of embryonic stem-like cells from mouse testis. The importance of our simple method will be clear when we compared it with other complex and time-consuming methods. Embryonic stem-like colonies with sharp border confirmed by real-time PCR, immunocytochemistry and flow cytometry assessments. Embryonic stem-like colonies were immunopositive for pluripotency markers. Transition of spermatogonia cells to embryonic stem-like cells was accompanied by extensive changes in gene expression. These changes included significant increase in pluripotency genes expression and significant decrease in germ cellspecific genes expression. Also, we proved the differentiation capacity of embryonic stem-like cells to neuroepithelial-like cells which were immunoreactive to Nestin and Neurofilament 68. Evaluation of genes expression during in vitro differentiation into neuroepithelial-like cells showed high-level expression of Nestin whether this gene approximately has no expression in undifferentiated embryonic stem-like cells. Also, expression of pluripotency genes has significantly decreased in neuroepithelial-like cells compared with embryonic stem-like cells. This study shows that embryonic stem-like cells derived from testis are capable to differentiate into neuroepitheliallike cells that may provide a cellular reservoir usable for neurodegenerative disorders.

Expression of Spermatogonial and Pluripotency Markers in Spermatogonial Stem Cells after Treatment with Different Culture Factors

“…In fact Oct4 has been shown to function as core transcription factor in maintain self-renewal of embryonic stem cells and SSCs. This data is similar to previous results that reported the differentiation of SSCs into different lineage specifically neural precursor under the define culture conditions (13,15,(32)(33). Mizrak et al mentioned that culture conditions support the differentiation of SSCs (14).…”

Silk nanofibrous electrospun scaffold enhances differentiation of embryonic stem like cells derived from testis in to mature neuron

“…Similar to previous reports these inducers can improve the neural differentiation in pluripotent stem cells population include: ES and ES Like cells. 18,19,23 Our date indicated that cells grown on Silk scaffold were significantly higher than monolayer cell group. Evaluation of cell culture seeded in silk scaffold confirmed that nanofibers structures can regulate cellular adherence, viability and proliferation.…”

“…Improved expression of mature neuron markers: Map2 and ß tubulin shows the functional promoting role of theses regents in neural maturation. Similar to previous reports these inducers can improve the neural differentiation in pluripotent stem cells population include: ES and ES Like cells …”

“…This study performed after obtaining animal ethical clearance from Mazandarn University of Medical Science. ES like cells were induced from neonatal mouse testis according our previously report . Briefly, testis were collected from Neonatal mouse using small insecure in lower part of the abdomen.…”

“…ES like cells were induced from neonatal mouse testis according our previously report. 23 Briefly, testis were collected from Neonatal mouse using small insecure in lower part of the abdomen. Testicular tissue was dissociated via a two-step mechanical and enzymatic Digestion.…”