ABSTRACT
In the past 10 years, adipose-derived stem cells (ADSCs) have been applied due to their pluripotency. Experimental tissues have been frequently obtained from mammals, including rabbits, mice and humans, but rarely from broilers, Gallus gallus domesticus. In the present study, ADSCs were obtained from 20-day-old broiler embryos. Primary ADSCs were sub-cultured to passage 37 in vitro. The surface markers of ADSCs, namely CD29, CD31, CD44, CD71 and CD73, were detected by reverse transcription polymerase chain reaction and immunofluorescence assays. The result indicated that CD29, CD44, CD71 and CD73 were expressed on the surface of cells at various passages, but not CD31. The growth curve of cells at the different passages had a typical sigmoidal shape. Furthermore, ADSCs were successfully induced to differentiate into osteoblasts, adipocytes and hepatocyte-like cells. The results denote that the ADSCs isolated from broilers have similar biological properties to those of ADSCs obtained from other animals. The present study provided a theoretical and experimental foundation for the use of poultry as a source of stem cells, and laid a foundation for adipose tissue engineering and strategies in regenerative medicine.
ABSTRACT
In vitro production of functional spermatids has special significance in the research of spermatogenesis and the treatment of male infertility. Primordial germ cells (PGCs) are the precursors of oocyte and sperm, which generate the totipotent cells. Studies have shown that PGCs have the potential ability to develop meiotic spermatids in vitro. Here we have shown that retinoic acid (RA) leads to PGC differentiation, and SCF can improve the efficiency of induction. We indicate an efficient approach to produce haploid spermatids from chicken PGCs in the presence of RA and stem cell factor (SCF). Real-time RT-PCR assays showed that RA and SCF induced a remarkable increase in expression of SYCP1, ACR, BOULE and DCM1 of meiotic germ cells and haploid germ cells, respectively. DNA content assays revealed that RA and SCF induced a remarkable increase of haploid cells. This study provides a theoretical basis and a great animal model for spermatogenesis study.
Subject(s)
Haploidy , Spermatids/cytology , Animals , Biomarkers/metabolism , Cell Shape/drug effects , Chick Embryo , DNA/metabolism , Female , Gene Expression Regulation, Developmental/drug effects , Male , Meiosis/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Spermatids/drug effects , Spermatids/metabolism , Stem Cell Factor/pharmacology , Tretinoin/pharmacologyABSTRACT
Mesenchymal stem cells derived from amniotic fluid have become one of the most potential stem cell source for cell-based therapy for the reason they can be harvested at low cost and without ethical problems. Here, we obtained amniotic fluid stem cells (AFSCs) from ovine amniotic fluid and studied the expansion capacity, cell markers expression, karyotype, and multilineage differentiation ability. In our work, AFSCs were subcultured to passage 62. The cell markers, CD29, CD44, CD73 and OCT4 which analyzed by RT-PCR were positive; CD44, CD73, CD90, CD105, NANOG, OCT4 analyzed by immunofluorescence and flow cytometry were also positive. The growth curves of different passages were all typically sigmoidal. The different passages cells took on a normal karyotype. In addition, AFSCs were successfully induced to differentiate into adipocytes, osteoblasts and chondrocytes. The results suggested that the AFSCs isolated from ovine maintained normal biological characteristics and their multilineage differentiation potential provides many potential applications in cell-based therapies and tissue engineering.
