The production of feline induced pluripotent stem cells (iPSCs) can solve the problems that are related with existing unstable supply and demand of eggs as well as ethical aspects about embryonic stem cell at the same time. On the basis of excellent proliferation, it is to facilitate the researches about human disease like FIV and Allergen at the level of cells, not experimental animals. But, a lot of advanced researches are lean too much towards on the transduction using DNA type virus that have the risk of tumorigenesis during reprogramming and on the mLIF-dependent culture condition for the production of feline iPSCs. This being so, this study shows the reprogramming results using Sendai virus vector that is RNA type virus and have no the footprint after transduction. In addition, the feline iPSCs were stably cultured in bFGF-dependent culture condition during the reprogramming step and culture step. In conclusion, we found the bFGF-dependent culture condition in feline iPSCs and suggested the approach using Sendai virus vector as an alternative for reprogramming without concern about tumorigenesis. These methods can be universally applicable to not only the researches about reconstruction and conservation of feline species, but also to a lot of deep studies related with iPSCs or LIF, bFGF to find new approaches.
The puφose of this study was to evaluate the role of EGF(Epidermal Growth Factor), EGFR(Epidermal Growth Factor Receptor), aFGF(acidic Fibroblast Growth Factor or FGF-1), bFGF(basic Fibroblast Growth Factor or FGF-2), FGFR(Fibroblast Growth Faαor Receptor), and VEGF(Vascular Endothelial Growth Factor) in the development of the human ameloblastomas For this study 9 subjects, diagnosed as ameloblastomas referred to the Dept. of Oral Path. College of Dentistry, Kyung Hee University, 2 subjeαs of normal oral mucosa with any inflammatory changes were used as experimental, control groups respectively. All the tissues ; experimental and control group were fixed in 10% neutral formalin solution and embedded in paraffm, serial tissue section were made 하1m in thickness and processed in the standard way for immunohistochemical method, using primary and secondary antibodies, for EGF(Antirabbit Ig G, rabbit kit at }:1oo dilution), EGFR(Antimouse Ig G, mouse kit at 1:100 dilution), aFGF(Antirabbit Ig G, rabbit kit at 1:100 dilution), bFGF(Antirabbit Ig G, rabbit kit at 1:100 dilution), FGFR(Antimouse Ig G, mouse kit at 1:100 dilution) , and VEGF(Antirabbit Ig G, rabbit kit at 1:100 dilution), all BioGenex U.S. A. made, followed by the Streptavidin - Horse Radish Peroxidase(InnoGenex, Human-avidin kit) appli때on , counter stained with Mayer’s hematoxylin stain method. And examined under the biologic microscope, graded -(no epithelial stain), +(weak or focal epithelial stain), ++(moderate or focal intensive epithelial stain), +++(intense generalized epithelial staining) for the epithelial, and connective 따sue component in ameloblastomas and in normal mucosal epithelium on each. Attained results as follows ; 1. EGF, EGFR, aFGF, bFGF, FGFR, and VEGF showed more intense stainability on experimental group compare t,。 that on the control group. 2. EGF, EGFR, aFGF, bFGF, FGFR, and VEGF showed more intense stainability in epithelial component and more intensely stained on the peripheral ce11s of the ameloblastomas. 3. EGF, EGFR, aFGF, bFGF, FGFR, and VEGF showed positive stainability on the stromal tissues but its level is lower compare to that on the epithelial components. Those results suggested that those growth factors take a role in development and progression on the amelob비las와tomas
까le purpose of this study was to evaluate the role of EGF(Epidermal Growth Factor) , EGFR(Epidemlal Growth Factor Receptor), aFGF(acidic Fibroblast Growth Factor, FGF-1), bFGF(basic Fibroblast Growth Factor, FGF-2), FGFR(Fibroblast Growth Factor Receptor), and VEGF(Vascular Endothelial Growth Factor) 띠 the development of the oral squamous cell carcinoma. For this study 6 subjects, diagnosed as squamous cell carcinoma refelTed to the Dept. of Oral Path. College of Dentistry, Kyung Hee University, 2 subjects of normal 이띠 mucosa with any inflammatolY changes were used as expelimental, control groups respectively. AlI the 디ssu es ; expe디me nta l and control group were fixed in 100;ú neutral fOlmalin solution and embeclded in paraffìn , seIial tissue section were made 511m in thickness ancl processecl in the standard way for immunohistochemical methocl, using primary ancl seconclalY antibodies, for EGF(Antirabbit Ig G, rabbit kit at 1:100 dilution), EGFR(Antimouse Ig G, mouse kit at 1:100 dilution), aFGF(Antirabbit Ig G, rabbit kit at 1:100 dilution), bFGF(Antirabbit Ig G, rabbit 써t at 1:100 dilution), FGFR(Antimouse Ig G, mouse kit at 1:100 dilution), ancl VEGF(Antirabbit Ig G, rabbit kit at 1:100 clilution), all BioGenex U.S.A. macle, followed by the Stre ptavidin - Horse Radish Peroxidase(InnoGenex, Human-avidin kit) application, counter stained with Mayer’s hematoxylin stain method. And examined under the biologic microscope, -(no epithelial stain), +(weak or focal epithelial stain), ++(mode rate or focal intensive epithelial stain), +++(intense generalized epithelial staining) for the epithelial, and connective tissue component in squamous cell carcinoma and in nomlal mucosal epithelium on each. Attained results as follows ; 1. It is noted that more intensed reactio n EGF, EGFR, aFGF, bFGF, FGFR, and VEGF on experimental group compare to that on the control group. 2. Increased reaction is noted on the tumor components compare to that in the stromal tissues. 3. Intensed reaction is noted on the basement membrane adjacent to cancer nest to EGF, EGFR, aFGF, bFGF, FGFR, and VEGF 4. It is noted that intensed positive reaction on cancer pearls, cancer components with hyperactivities, in cancer nest. And at the peIiphelY of cancer nest, diffuse moclerate reaction to EGF, EGFR, aFGF, bFGF, FGFR, and VEGF is notecl This results suggest that EGF, EGFR, aFGF, bFGF, FGFR, and VEGF mJy be effectecl to the growth ancl clevelopment of the squamous cell carcinoma.
The purpose of this study was to evaluate the role of EGF(Epidermal Growth Factor), EGFR(Epidermal Growth Factor Receptor), aFGF(acidic Fibroblast Growth Factor, FGF-1), bFGF(basic Fibroblast Growth Factor, FGF-2), FGFR(Fibroblast Growth Factor Receptor) in the development of radicular cyst. For this study 37 subjects, diagnosed as radicular cysts. referred to the Dept. of Oral Path. College of Dentistry, Kyung Hee University, were used as experimental group. And for control group, 2 subjects of normal oral epithelium without any inflammatory changes were used. All the tissues; experimental and control group were neutral formation fixed and paraffin embedded. serial tissue section were made at 5㎛ and processed in the standard way for immunohistochemical method, using primary antibodies against, EGF(Antirabbit Ig G at 1:100 dilution), EGFR(Antimouse Ig G at 1:100 dilution), aFGF(Antirabbit Ig G, rabbit kit at 1:100 dilution), bFGF(Antirabbit Ig G, mouse kit at 1:100 dilution), FGFR(Antimouse Ig G mouse kit at 1:100 dilution), all BioGenex U.S.A. made except EGFR(Chemicon U.S.A.) followed by the Streptavidin - Horse Radish Peroxidase (InnoGenex Human-avidin kit) application, counter stained with Meyer's hematoxylin stain method. And examined under microscope, graded 0(no epithelial stain), +(weak or focal epithelial stain), ++(moderate or focal intensive epithelial stain), +++(intense generalized epithelial staining) for the epithelium, and connective tissue of cyst wall.
1. EGF, EGFR, aFGF, bFGF, FGFR showed more intense staining on radicular cysts compare to that on the normal
mucosa.
2. EGF, EGFR, aFGF, bFGF, FGFR stained in mucosa, submucosa of the control group and also stained on the lining
epithelium, connective tissues of cyst wall in the experimental group.
EGF, EGFR, aFGF, bFGF, FGFR take a part in the development of the radicular cyst.
Multipotent perivascular stem cells (PVCs) have gained much attention as an alternative source for cell based regenerative medicine in recent years. Due to their rarity in human tissues, developing methods to efficiently isolate and expand PVCs from various fetal and adult tissues is necessary to obtain a clinically relevant number of cells that maintain progenitor potency. Here, we report on a non-enzymatic isolation (NE) method of PVCs from human umbilical cords (HUCs) and compare its efficiency with the conventional collagenase treatment method (CT) in terms of proliferation and immunophenotypes. The cells isolated by NE displyed acceptable surface marker profile of PVCs and showed multilineage (osteogenic, chondrogenic, and adipogenic) differentiation potential. While both methods provided similar levels or patterns of proliferation and immunophenotypes, PVCs by NE retained a higher level of CD146(+) frequency compared to that of CT over passage. Furthermore, we have investigated potentials of various exogenous factors to promote proliferation of HUCPVCs in vitro. Among these factors, supplementation of basic fibroblast growth factor (bFGF) provided the optimal condition to significantly enhance the proliferation rate of HUCPVC and increased a proportion of stage-specific antigen-4 (SSEA-4) positive subset. Collectively, our study suggests that NE method with bFGF supplementation offers an alternative way to obtain sufficient numbers of HUCPVCs with higher number of primitive SSEA-4(+) subpopulation that are applicable in therapeutic doses for regenerative medicine.
Pluripotent cells are categorized as either "naive" or "primed" based upon their pluripotent status. According to previous studies, embryonic stem cells and embryonic germ cells are identified as naive pluripotent stem cells and epiblast stem cells are identified as primed pluripotent stem cells. In a permissive species such as the mouse, naive and primed pluripotent stem cells can be derived from embryos without genetic manipulations. In non-permissive species such as humans and pigs, primed pluripotent cells are only established from embryos. However, previous studies have shown that the embryonic germ cells of non-permissive species share similar morphology and features with naive pluripotent cells. For these reasons porcine embryonic germ cells (pEGCs) may provide a useful cell source for comparative studies on naive pluripotent cells in non-permissive species. In this study, we attempted to establish and characterize porcine embryonic germ cells. Consequently, an embryonic germ cell line was derived from the genital ridges of a porcine dpc 30 fetus in media containing bFGF. This cell line displayed a dome-shaped colony morphology. The cell line was maintained in a stable condition over an extended time period and was able to differentiate into the three germ layers in vitro. Pluripotency markers such as OCT4, SOX2, NANOG and SSEA4 were expressed in these pEGCs. Similar with pESCs, Mek/Erk signaling pathway were activated by bFGF in the cultured pEGCs. In conclusion, we were able to successfully derive embryonic germ cells from genital ridges of a porcine fetus. Unlikely naive pluripotent cells such as mESCs, pluripotency of pEGCs were regulated by Mek/Erk signaling pathway activated by bFGF. This cell line could potentially be used as naive pluripotent cell source for comparative study with porcine embryonic stem cells and other pluripotent cell lines. As porcine pluripotent cells, pEGCs could be useful candidates for preliminary studies of human disease as well as a source for generating transgenic animals.