CD105, a polyclonal rabbit antibody (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), was used as a marker for rat stem cells.
McKay and colleague Robert Tsai, also of NINDS, first found the gene in cultured rat stem cells.
In a 2010 Cell paper, Nakauchi's team described growing rat pancreases, using rat stem cells, in apancreatic mouse embryos.
Mutant mice are traditionally crafted by disabling genes in embryonic stem cells, but no one could get rat stem cells with incapacitated genes to survive in the laboratory.
This result is consistent with the previous study, which shows that TRPC5 predominantly expresses in the CNS and is involved in proliferation of embryonic rat stem cells [23].
We suggest, that a reporter gene construct, driven by the Nanog promoter in transgenic rats, similar to what was published for mouse [24], would help to enhance the selection process of germline-competent ES cells during the derivation from rat preimplantation embryos and induced pluripotent rat stem cells.
The rat stem cells grew in the special mouse environment in the absent of mouse pancreas.
When the mice matured to adulthood, they developed a rat pancreas from the injected rat stem cells.
Briefly, LDBM cells were cultured for 2 days in Iscove's Modified Dulbecco's Medium (Invitrogen) supplemented with 10% fetal calf serum (FCS, HyClone, Logan, UT), 2% penicillin and streptomycin (P/S), and 100 ng/ml each of recombinant rat stem cell factor (SCF), megakaryocyte growth and development factor (MGDF) and granulocyte colony-stimulating factor (G-CSF) (all from Amgen, Thousand Oaks, CA).
Uterine-like tissues were then regenerated and maintained in vitro for up to 10 d through decellularized uterine matrix (DUM) reseeding with adult and neonatal rat uterine cells and rat mesenchymal stem cells followed by aortic perfusion in a bioreactor.
