• Anatomy and Cell Biology
• /
• v.56 no.1
• /
• pp.25-31
• /
• 2023

Johannes Nathanael Lieberkühn was a prodigious anatomist whose meticulous experiments and precise detailing helped in comprehending the microscopic anatomy of digestive system during early part of eighteenth century. Notably, his inventions in the field of microscopy aptly complemented his quest for anatomical knowledge at microscopic level. He designed a reflector (Lieberkühn reflector) which enhanced the amount of focussed light leading to bright illumination of tissue specimen. He invented the solar microscope which provided excellent resolution of minute anatomical details. Lieberkühn discovered the digestive juice secreting tubular glands (glands of Lieberkühn) present at the base of intestinal villi producing epithelial invaginations (crypts of Lieberkühn). He also described the intricate juxtaposition of blood vessels in relation to a single intestinal villi. Moreover, through empirically designed experimental set up, Lieberkühn was able to demonstrate the flow of lymph from intestinal villi to collecting lymphatic vessels. Also, his grandiose collection of laboratory specimens involving vascular anatomy are a testimony of his untiring efforts in academia. His contributions were seminal in comprehending the anatomy of digestive system and paved the way for future revelations. His work unveiled the enormous scope of microanatomy in medical science and catalysed the advent of histological staining methods a century later.

• Molecules and Cells
• /
• v.41 no.4
• /
• pp.290-300
• /
• 2018

Using an in vitro model of intestinal organoids derived from intestinal crypts, we examined effects of indole-3-carbinol (I3C), a phytochemical that has anticancer and aryl hydrocarbon receptor (AhR)-activating abilities and thus is sold as a dietary supplement, on the development of intestinal organoids and investigated the underlying mechanisms. I3C inhibited the in vitro development of mouse intestinal organoids. Addition of ${\alpha}$-naphthoflavone, an AhR antagonist or AhR siRNA transfection, suppressed I3C function, suggesting that I3C-mediated interference with organoid development is AhR-dependent. I3C increased the expression of Muc2 and lysozyme, lineage-specific genes for goblet cells and Paneth cells, respectively, but inhibits the expression of IAP, a marker gene for enterocytes. In the intestines of mice treated with I3C, the number of goblet cells was reduced, but the number of Paneth cells and the depth and length of crypts and villi were not changed. I3C increased the level of active nonphosphorylated ${\beta}$-catenin, but suppressed the Notch signal. As a result, expression of Hes1, a Notch target gene and a transcriptional repressor that plays a key role in enterocyte differentiation, was reduced, whereas expression of Math1, involved in the differentiation of secretory lineages, was increased. These results provide direct evidence for the role of AhR in the regulation of the development of intestinal stem cells and indicate that such regulation is likely mediated by regulation of Wnt and Notch signals.

• Journal of Animal Science and Technology
• /
• v.64 no.6
• /
• pp.1105-1116
• /
• 2022

Recently, we reported the robust in vitro three-dimensional (3D) expansion of intestinal organoids derived from adult bovine (> 24 months) samples. The present study aimed to establish an in vitro 3D system for the cultivation of intestinal organoids derived from growing cattle (12 months old) for practical use as a potential alternative to in vivo systems for various purposes. However, very few studies on the functional characterization and 3D expansion of adult stem cells from livestock species compared to those from other species are available. In this study, intestinal crypts, including intestinal stem cells, from the small intestines (ileum and jejunum) of growing cattle were isolated and long-term 3D cultures were successfully established using a scaffold-based method. Furthermore, we generated an apical-out intestinal organoid derived from growing cattle. Interestingly, intestinal organoids derived from the ileum, but not the jejunum, could be expanded without losing the ability to recapitulate crypts, and these organoids specifically expressed several specific markers of intestinal stem cells and the intestinal epithelium. Furthermore, these organoids exhibited key functionality with regard to high permeability for compounds up to 4 kDa in size (e.g., fluorescein isothiocyanate [FITC]-dextran), indicating that apical-out intestinal organoids are better than other models. Collectively, these results indicate the establishment of growing cattle-derived intestinal organoids and subsequent generation of apical-out intestinal organoids. These organoids may be valuable tools and potential alternatives to in vivo systems for examining host-pathogen interactions involving epithelial cells, such as enteric virus infection and nutrient absorption, and may be used for various purposes.

• Food Science and Biotechnology
• /
• v.15 no.6
• /
• pp.931-936
• /
• 2006

This study sought to investigate any additive or interactive effects that soy isoflavones may have on the ecosystem of the gut, which is influenced by fructooligosaccharide (FOS) in colon-cancer model rats. Male Sprague-Dawley rats treated with 1,2-dimethylhydrazine were given experimental diets containing 0, 3, 6, or 9% FOS with or without 0.1% soy isoflavone for 12 weeks. In addition to the effects of FOS dosage on the gut ecosystem, dietary supplementation with soy isoflavone reduced the number of colonic aberrant crypts (ACs). The fecal weight, fecal pH, and gut transit time significantly decreased in a dose-dependent manner in rats fed FOS and the fecal concentration of bifidobacteria was higher in rats fed FOS than in control rats. The fecal output of total short-chain fatty acids, acetate, and propionate was significantly increased by the presence of FOS and was negatively correlated with the number of ACs, whereas the fecal output of butyrate showed no significant correlation with FOS dosage. The addition of soy isoflavone to the diet did not result in any significant differences in gut ecosystem parameters. Therefore, we conclude that the suppressive effect of soy isoflavone on ACs was not associated with the intestinal ecosystem, which was significantly altered by the dosage of FOS.

• Radiation Oncology Journal
• /
• v.19 no.3
• /
• pp.259-264
• /
• 2001

Purpose : We investigated the temporal alterations of apoptosis and mitotic death following irradiation in the rat's small intestinal crypts. Materials and methods : Male Sprague-Dawley rats were irradiated 2 Gy by 6 MV linear accelerator and sacrified at 2, 4, 8, 24, 48 hours after irradiation. The mean numbers of the apoptotic cells and mitotic cells per their small intestinal crypts were measured in the unirradiated control and irradiated groups. To compare with H & E staining, ISEL (In Situ End Labelling) were peformed in the group having the highest apoptotic count. Results : The mean number of the apoptosis per crypt in the control group was 0.14 and those at 2, 4, 8, 24, 48 hours after irradiation were 1.43, 3.19, 1.15, 0.26, 0.17, respectively. So the apoptosis development was increased upto 4 hours and then normalized around 24 hours following irradiation. The mean number of the mitotic cells per crypt in the control group was 1.29 and those at 2, 4, 8, 24, 48 hours after irradiation were 0.56, 0.47, 0.23, 0.65, 1.19, respectively. The mitotic cell counts following irradiation was decreased to 8 hours and recovered to the normal level about 48 hours. So the increment of apoptotic cell count was occurred earlier and more remarkable than the decrement of mitotic cell count after irradiation. According to the staining time, false positivity was found in the ISEL staining. Conclusions : The cell death in the small intestinal crypt developed by acute radiation damage was usually decreased to the normal level within $24\~48\;hours$ after irradiation and the apoptosis was thought to be more important process than the mitotic death.

• Korean Journal of Veterinary Research
• /
• v.34 no.3
• /
• pp.537-547
• /
• 1994

Ninety seven cases of histopathologically diagnosed spontaneous canine parvovirus enteritis(CPE) were studied gross pathologically, histopathologically, immunohistochemically, to investigate histopathological types of small intestinal lesions, and antigen distributions in each pattern related to the infected age. And also, reliability of histopathological method in diagnosis of CPE was inspected with immunohistochemistry. The results were as follows : 1. Age-related occurring ratio in histopathologically diagnosed CPE was 53.6% in 4-8 weeks, 26.8% in 9-15 weeks, 8.25 in 16-19 weeks and 11.3% in 20-45 weeks of the clog age. 2. In histopathologic classification based on patterns of villi/crypts lesions of small intestine(jejunum), the ratio of A type (initial phase of necrosis of crypt epithelia, desquamated epithelial cells in the dilated lumen of the crypt) was 20.6%; the ratio of B type(middle phase of atrophy and fission of the villi, collapse of the mucosa, loss of normal crypt structure) was 62.9%, and C type(regenerative phase of the crypt architecture) was 16.5%. 3. The ratio of A, B, C type in 4-8 weeks old, respectively, was 23.5%, 61.5%, 15.4%; in 9-15 weeks old was 19.2%, 65.4%, 15.3% in 16-19 weeks old was 25.0%, 75.0%, 0.0%; and in 20-45 weeks old was 9.0%, 54.5%, 36.4%. 4. The antigen distribution in the nuclei of the crypt epithelial cells was higher than of the cytoplasm and numerous desquamated epithelial cells in dialated crypts in A type; The antigen cytoplasm and numerous desquamated epithelial cells in dialated crypts in A type; The antigen distribution in the nuclei of the collapsed crypt epithelial cells was not higher than that of the cytoplasm, crypts were lined by and filled with released viral antigens from the destructed epithelial cells in B type; and its distribution was also higher than in the epithelial cells adjacent to the tips of the villi, but it was not reacted in the regenerative crypt epithelial cells in C type. 5. Immunohistochemically detected antigen ratio in the small intestine of histopathologically diagnosed CPE was 94.6%, and this result indicates that histopathological diagnosis is very reliable method in diagnosis of CPE.

• Journal of Animal Reproduction and Biotechnology
• /
• v.39 no.1
• /
• pp.2-11
• /
• 2024

Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

• Journal of Animal Reproduction and Biotechnology
• /
• v.37 no.2
• /
• pp.136-143
• /
• 2022

Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1 µM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.

• Korean Journal of Veterinary Research
• /
• v.37 no.1
• /
• pp.161-166
• /
• 1997

From January of 1991 to December of 1992, 42 chickens collected from 21 poultry farms and also diagnosed as necrotic enteritis(NE) was examined clinical signs, gross and histopathological findings. Main clinical signs were characterized by decreased appetite, mild to severe depression, reductance to move, ruffled feathers, greenish to yellow-browinish diarrhea sometimes including blood. As progressed, diseased chickens showed feces mixed with necrotic debris which detached from the intestinal mucosa and mostly resulted in the death. In chronic cases, there were dirty feathers around cloaca due to diarrhea and notably retarded growth. Principle gross lesions were usually confined to the jejunum and ileum, especially toward the lower part of Meckel's diverticulum. The part of small intestine was frequently distended with gas, and also showed mucosal congestion and hemorrhages with varying degrees. Sometimes, the intestinal mucosa was thickened, and also covered with fibronecrotic psuedomembrane. In addition, there were focal necrosis and severely multifocal ulcreation in the mucosa of small intestine. Major histopathological findings included villous necrosis and erosion of the small intestine covering with lots of bacterial colonies, inflammatory cell infiltration in the lamina propria, and dilatation and hyperplasia of crypts. Luminal exudate contained bacterial colonies, fibrin, erythrocytes, and desquamated epithelium. Thirteen(61.9%) out of 21 NE-occurring farms were complicated with intestinal coccidiosis.

• Parasites, Hosts and Diseases
• /
• v.41 no.1
• /
• pp.63-67
• /
• 2003

A 69-year-old Korean man was admitted to emergency room with complaints of abdominal pain, vomiting, and diarrhea, Laboratory tests revealed eosinophilia, anemia, hypoproteinemia, and hyponatremia. The gastric mucosa showed whitish mottled and slightly elevated lesions on the body angle of antrum. Microscopically, chronic gastritis with incomplete intestinal metaplasia was observed. Many adult worms, larvae, and eggs in cross sections were located in the crypts. Furthermore, the filariform larvae of Strongyloides stercoralis with a notched tail were detected through the culture.