مقالات پذیرفته شده در سومین کنگره بین المللی زیست پزشکی
Effects of osteocalcin in proliferation and differentiation of spermatogonial cells cocultured with somatic cells
Effects of osteocalcin in proliferation and differentiation of spermatogonial cells cocultured with somatic cells
Tayebeh Rastegar,1,*Somayeh Solhjoo,2Heidar Toolee,3
1. tehran university of medical sciences 2. kerman university of medical sciences 3. Shahroud university of medical sciences
Introduction: Spermatogonial cells (SCs) are key cells for spermatogenesis. These cells are
affected by paracrine signals originated from nearby somatic cells, among them
Leydig cells have receptors for osteocalcin, a hormone known for exerting positive
roles in the promotion of spermatogenesis. The aim of this study was to evaluate
roles for osteocalcin on SCs proliferative and differentiation features after coculture
with Leydig cells.
Methods: SCs and Leydig cells were isolated from neonate NMRI offspring
mice and adult NMRI mice, respectively. SCs population were then enriched in a
differential attachment technique and assessed for morphological features and
identity. Then, SCs were cocultured with Leydig cells and incubated with
osteocalcin for 4 weeks. Evaluation of proliferation and differentiation‐related
factors were surveyed using immunocytochemistry (ICC), Western blot, and
quantitative real‐time polymerase chain reaction (PCR). Finally, the rate of
testosterone release to the culture media was measured at the end of 4th week.
Results: Morphological and flow cytometry results showed that the SCs were the population
of cells able to form colonies and to express ID4, α6‐, and β1‐integrin markers,
respectively. Leydig cells were also able to express Gprc6α as a specific marker for
the cells. Incubation of SCs/Leydig coculture with osteocalcin has resulted in an
increase in the rate of expressions for differentiation‐related markers. Levels of
testosterone in the culture media of SCs/Leydig was positively influenced by
osteocalcin.
Conclusion: It could be concluded that osteocalcin acts as a positive inducer of SCs
in coculture with Leydig cells probably through stimulation of testosterone release
from Leydig cells and associated signaling.