A cat who is a 15-year-old and spayed female visited an animal clinic with severe coughing symptoms. Since the cat’s coughing symptoms had worsened from the age of 10 and X-rays showed a bronchial pattern in the lungs, it was diagnosed as Chronic Obstructive Pulmonary Disease (COPD). She received three injections of stem cells isolated from the amniotic membrane on days 0, 7, and 23. Although there was no improvement in the clinical findings on the x-ray, the number of coughing was significantly reduced. In addition, even after long-term follow-up post treatment for a month, she was stable with almost no coughing.

Three different cats who had chronic kidney disease (CKD) were treated for more than one month with fluid therapy in an animal clinic. Although this long-term treatment and hospitalization, there was no clinical improvement in clinical signs as well as serum biochemical indexes including blood urea nitrogen (BUN), creatinine (CREA), and phosphate (PHOS). All cases were then injected three times with allogeneic stem cells through an intravenous route for treatment on Day 0, 7, and 14 or 30. On the same day, clinical observation and blood tests for serum biochemistry were conducted together. Upon administrating stem cells to the CKD cats, clinical conditions and the indexes of BUN and CREA were clinically improved within normal ranges. Additionally, one of the cats who had the renal cysts presented clinical improvement with showing decreased cysts size than before.

Amniotic membrane stem cells are considered as a good alternative to embryonic stem cells, but their use in clinical studies is still not common. Here, exosomes from canine amniotic membrane mesenchymal stem cells (cAmMSCexo) were used for dog sperm cryopreservation. Upon cryopreserved straws using cryoprotectant containing 0, 0.5, 1, or 2 μg/mL of cAmMSC-exo were thawed, motility and membrane integrity were analyzed. However, results showed no significant differences between the groups. We concluded that cAmMSC-exo with lower than 2 µg/mL have no effects on sperm cryopreservation, and further studies to get higher concentrations of cAmMSC-exo should be conducted for clinical application.

The present study was conducted to investigate gelatinolytic activities in HAM and to determine whether there are any changes in gelatinolytic activity profiles when the cells are cultured in hepatogenic medium. Placenta was obtained during caesarean section of the volunteers, with informed consent. HAM were isolated from amniotic membrane using collagenase type A HAM were cultured in hepatogenic medium for 3 weeks and the conditioned media were obtained at day 7, 14 and 21. The zymographic pattern of gelatinolytic activity of the HAM did not undergo a change during passages. When the HAM were cultured in a fibronectin-coated dishes in a hepatogenic medium, there was no significant difference of the gelatinase pattern between before and after culture. However, when bFGF was added to the culture, a dramatic increase of 62kDa and 59kDa gelatinases was observed. Interestingly, when ITS instead of FN was present, HAM-conditioned medium also showed a similar increase of both gelatinases. Immunoblotting analysis demonstrated that both 62kDa and 59kDa gelatinases were the active form of MMP-2 resulting from the turnover of MMP-2 proform. Futher study will be necessary to determine the relationship between bFGF and active MMP-2 during hepatogenesis of HAM.

This study was designed to compare re-epithelization rate/feature of conjunctival epithelial cell on amniotic membrane and anatomical difference between normal amniotic membrane and de-epithelial amniotic membrane. Human conjunctival tissue obtained while cataract operation were used in this study. Primary cultured human conjunctival epithelial cell and fibroblast were incubated on intact amniotic membrane and de-epithelialized amniotic membrane. After incubation for 3, 5 and 7 days, re-epithelization rate was analyzed using cell tracker and microscopic feature was examined using methylene blue and hematoxylin- eosin stain. For cell marker analysis, cytokeratin 14 and vimentin antibody immunofluorescence stain were used. Clearance rate of epithelial cells on amniotic membrane using trypsin was better than using 25% alcohol. The results of reepithelization rate using cell tracker, HaCaT, conjunctival firbroblast, and epithelial cell were rapidly incubated on deepithelialized amniotic membrane. Wound healing and re-epithelization were more rapidly induced on de-epithelialized amniotic membrane on permanent amniotic membrane graft.

Human eyelid adipose-derived stem cells (hEAs) and amniotic mesenchymal stem cells (hAMs) are very valuable sources for the cell therapeutics. Both types of cells have a great proliferating ability in vitro and a multipotency to differentiate into adipocytes, osteoblasts and chondrocytes. In the present study, we evaluated their stem cell characteristics after long-time cryopreservation for 6, 12 and 24 months. When frozen-thawed cells were cultivated in vitro, their cumulative cell number and doubling time were similar to freshly prepared cells. Also they expressed stem cell-related genes of SCF, NANOG, OCT4, and TERT, ectoderm-related genes of NCAM and FGF5, mesoderm/endoderm-related genes of CK18 and VIM, and immune-related genes of HLA-ABC and 2M. Following differentiation culture in appropriate culture media for 2-3 weeks, both types of cells exhibited well differentiation into adipocyte, osteoblast, and chondrocyte, as revealed by adipogenic, osteogenic or chondrogenic-specific staining and related genes, respectively. In conclusion, even after long-term storage hEAs and hAMs could maintain their stem cell characteristics, suggesting that they might be suitable for clinical application based on stem cell therapy.

간질환 환자의 대부분은 간 조직 손상으로 인해 간세포의 재생 능력이 감소한다. 간세포 이식은 이러한 간질환을 치료하는데 있어 혁신적인 방법으로 대두되고 있으나, 여전히 많은 의문과 문제점이 제기되고 있다. 사람의 양막으로부터 얻은 줄기 세포를 이용하여 간세포 분화를 위한 최적의 조건을 알아 보고자 하였다. 세포내 알부민에 대한 면역 화학적 방법, 세포내 글리코겐의 특이 염색법, 세포의 형태적 변화 연구 방법 등을 이용하여 여러가지 배양 조건을 조사한 결