ABSTRACT
In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.
Subject(s)
Humans , Infant , Infant, Newborn , Acrosin , Activins , Alkaline Phosphatase , Bone Morphogenetic Protein 4 , Dolichos , Follicle Stimulating Hormone , In Vitro Techniques , Spermatogenesis , Stem Cells , Swine , Testis , Testosterone , Tretinoin , Up-RegulationABSTRACT
We performed the transforming nitrate test mainly with penicillium brevi-compactum and other fungi. The strain of P. brevi-compactum and all the other fungi used for this test were isolated from patients with cervix uteri carcinoma, patients with gynopathy but no cancer, and the normal woman controls in Luoyang county of Shaanxi and in Wufeng county of Hubei respectively, which are the highest prevalance areas of cervix uteri carcinoma in China. The detection rate of P. brevicompactum is higher 72.73% (65/77) in Luoyang; 67.65 (23/34) in Wufeng. The detection rate of yeast and Candida is low. Though the detection rate of certain tested fungi such as A. serreus, A. candidus, P. cyelopium and Rhodotorula rubra is higher, these fungi are seldom found in the isolates from cervix uteri. Thus, these fungi might be no important significance in etiology of cervix uteri carcinoma. Because nitriamine is an important carcinogen and the effect of transforming nitrate to nitrite plays a more important role in the synthesis of nitriamine, P. brevi-compactum is the dominant strain of fungi isolated from the patient with cervix uteri carcinoma in this investigation. The higher detection rate of the treated P. brevi-compactum shows that this fungus plays a role in the etiology or incidence of cervis uteri carcinoma. To solve this difficult and complex problem of the role that fungal agent okay in the etiology of carcinoma further studies must be done.