To investigate the secretion of protoporphyrin IX (PpIX), superficial eggshell pigment, from shell gland cells of Japanese quail, the epithelial cells of the gland were collected and isolated for cultivation in vitro....To investigate the secretion of protoporphyrin IX (PpIX), superficial eggshell pigment, from shell gland cells of Japanese quail, the epithelial cells of the gland were collected and isolated for cultivation in vitro. An analysis of a peak for PpIX in the cells was performed using a fluorescence microplate reader. The measurement showed that PpIX has a peak of excitation wavelength at 410 nm and emission wavelength at 606 nm in the culture medium (HamF12 + 4% HCl). Volumes of PpIX in the medium after 4 hour culture of the cells were measured with a microplate reader using filter set of excitation wavelength 400/30nm and emission wavelength 620/40nm. However the cells did not secrete significantly PpIX during 4 hour incubation in this culture system, addition of quail plasma to the medium resulted in significantly higher secretion. A cultivation system in this study is able to use for the study on the mechanism of the secretion of eggshell pigment, PpIX from Japanese quail shell gland epithelial cells.展开更多
[ Objective] To develop in-vitro culture methods of yak endometrial gland epithelial cells. [Method] The gland epithelial cells were isolated from yak endometrium by explant culture method and digestion culture method...[ Objective] To develop in-vitro culture methods of yak endometrial gland epithelial cells. [Method] The gland epithelial cells were isolated from yak endometrium by explant culture method and digestion culture method, respectively. [ Result] In the first method, the cells isolated from the endometrium explant could merge into monolayer after 8-d culture, and they could be purified by gradation digestion with trypsin. In the second method, the endometrium explant were first digested by collagenase II by incubation at 37℃ for 2.5 h and then further digested in fresh 2 g/L colla- genase II for another 2.5 h. The cell suspension was leached through 74 pm filter and centrifuged at 400 r/min for 5 min. Then the cell pellet was re-suspended, followed by natural sedimentation to collect purified gland epithelial cells. The isolated cells were cytokeratin-positive as detected by immunocytochemical staining, and the positive rate could reach 95%. [Conclusion] The yak endometrial gland epithelial cells can be isolated and purified by both the explant culture method and digestion culture method.展开更多
Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To deter...Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C.展开更多
To investigate factors involved in the secretion of protoporphyrin IX (PpIX), a superficial eggshell pigment, from shell gland epithelial cells of Japanese quail, we cultured cells in Ham’s F12 medium with calcium ch...To investigate factors involved in the secretion of protoporphyrin IX (PpIX), a superficial eggshell pigment, from shell gland epithelial cells of Japanese quail, we cultured cells in Ham’s F12 medium with calcium chloride and quail plasma. The addition of hormones (prostaglandin F2α, progesterone, estradiol-17β) to the medium did not change the PpIX concentration in the culture supernatant, but changing the calcium chloride (CaCl2) concentration did: a lower concentration of CaCl2 led to a higher PpIX concentration;0 mM CaCl2 enhanced the secretion of PpIX from epithelial cells prepared at 5 or 7 mM CaCl2. The result suggests that a drop in concentration of CaCl2 mimics the end of shell calcification and stimulates rapid secretion of PpIX in vivo. Bovine serum albumin was almost as effective as quail plasma for PpIX secretion in culture, and would facilitate further study of the mechanism of PpIX secretion.展开更多
Craniopharynigoma samples were collected from 36 patients. Out of the 36 samples, 29 achieved successful sub-culturing, with a success rate of 80.6%. Immunohistochemistry staining showed that cytokeratin-7 was positiv...Craniopharynigoma samples were collected from 36 patients. Out of the 36 samples, 29 achieved successful sub-culturing, with a success rate of 80.6%. Immunohistochemistry staining showed that cytokeratin-7 was positively expressed in the cytomembrane and cytoplasm of craniopharyngioma cells at 6-8 passages, confirming that all cultured cells were squamous epithelial cells. The doubling time of craniopharyngioma cells was 3 days, as confirmed by the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. In this study, craniopharyngioma cells cultured in vitro were established; however, establishment of immortalized craniopharyngioma cell lines requires further research.展开更多
文摘To investigate the secretion of protoporphyrin IX (PpIX), superficial eggshell pigment, from shell gland cells of Japanese quail, the epithelial cells of the gland were collected and isolated for cultivation in vitro. An analysis of a peak for PpIX in the cells was performed using a fluorescence microplate reader. The measurement showed that PpIX has a peak of excitation wavelength at 410 nm and emission wavelength at 606 nm in the culture medium (HamF12 + 4% HCl). Volumes of PpIX in the medium after 4 hour culture of the cells were measured with a microplate reader using filter set of excitation wavelength 400/30nm and emission wavelength 620/40nm. However the cells did not secrete significantly PpIX during 4 hour incubation in this culture system, addition of quail plasma to the medium resulted in significantly higher secretion. A cultivation system in this study is able to use for the study on the mechanism of the secretion of eggshell pigment, PpIX from Japanese quail shell gland epithelial cells.
基金supported by the Key Project of Chinese Ministry of Education (210216)the Third Phase Construction Fee for High-Level Personnel of 211 Project (SZRC-211-05)
文摘[ Objective] To develop in-vitro culture methods of yak endometrial gland epithelial cells. [Method] The gland epithelial cells were isolated from yak endometrium by explant culture method and digestion culture method, respectively. [ Result] In the first method, the cells isolated from the endometrium explant could merge into monolayer after 8-d culture, and they could be purified by gradation digestion with trypsin. In the second method, the endometrium explant were first digested by collagenase II by incubation at 37℃ for 2.5 h and then further digested in fresh 2 g/L colla- genase II for another 2.5 h. The cell suspension was leached through 74 pm filter and centrifuged at 400 r/min for 5 min. Then the cell pellet was re-suspended, followed by natural sedimentation to collect purified gland epithelial cells. The isolated cells were cytokeratin-positive as detected by immunocytochemical staining, and the positive rate could reach 95%. [Conclusion] The yak endometrial gland epithelial cells can be isolated and purified by both the explant culture method and digestion culture method.
基金Supported by Doctoral Initial Fund of Ludong University (No.43304)
文摘Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C.
文摘To investigate factors involved in the secretion of protoporphyrin IX (PpIX), a superficial eggshell pigment, from shell gland epithelial cells of Japanese quail, we cultured cells in Ham’s F12 medium with calcium chloride and quail plasma. The addition of hormones (prostaglandin F2α, progesterone, estradiol-17β) to the medium did not change the PpIX concentration in the culture supernatant, but changing the calcium chloride (CaCl2) concentration did: a lower concentration of CaCl2 led to a higher PpIX concentration;0 mM CaCl2 enhanced the secretion of PpIX from epithelial cells prepared at 5 or 7 mM CaCl2. The result suggests that a drop in concentration of CaCl2 mimics the end of shell calcification and stimulates rapid secretion of PpIX in vivo. Bovine serum albumin was almost as effective as quail plasma for PpIX secretion in culture, and would facilitate further study of the mechanism of PpIX secretion.
基金supported by the National Natural Science Foundation of China, No. 30872646 and 30973082
文摘Craniopharynigoma samples were collected from 36 patients. Out of the 36 samples, 29 achieved successful sub-culturing, with a success rate of 80.6%. Immunohistochemistry staining showed that cytokeratin-7 was positively expressed in the cytomembrane and cytoplasm of craniopharyngioma cells at 6-8 passages, confirming that all cultured cells were squamous epithelial cells. The doubling time of craniopharyngioma cells was 3 days, as confirmed by the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay. In this study, craniopharyngioma cells cultured in vitro were established; however, establishment of immortalized craniopharyngioma cell lines requires further research.
