Use of icariin in inducting dry cell body in-vitro directional differentiation
A technology of icariin, stem cells, applied in the field of cell biology and pharmacology
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Embodiment 1
[0021] Icaritin-induced differentiation of embryonic stem cells (ES cells) into neural cells in vitro
[0022] 1. ES cell support culture:
[0023] ES cell D3 cell line (ES-D3 cell) in the basal medium (containing high glucose DMEM, non-essential amino acids, 10% fetal bovine serum, leukemia inhibitory factor), press 1 × 10 6 Cells / ml were inoculated on feeder cells (mouse fibroblasts), and subcultured for 2 to 3 days.
[0024] 2. Culture of icariin-induced ES cells to differentiate into neurons:
[0025] ES-D3 cells were digested with trypsin, and made into a single-cell suspension with differentiation medium (containing high-sugar DMEM, non-essential amino acids, 10% fetal bovine serum, and no leukemia inhibitory factor), at about 600 cells / 30 μL Inoculate on the inner surface of the lid of the petri dish, turn the lid over to form a hanging drop, and culture it in an incubator for 3 days. Thereafter, the hanging drops that had formed embryoid bodies were transferred to a...
Embodiment 2
[0026] Example 2 Use of icariin to induce directed differentiation of ES cells into nerve cells in drug efficacy evaluation If the embryoid body obtained in Example 1 can appear nerve-like cells (the length of the cell process is more than 5 times the diameter of the cell body) ), then use it as an indicator of directed differentiation into nerve cells, calculate the ratio of embryoid bodies that differentiated into neural-like cells to the total number of embryoid bodies, and make the dose-effect and sum of the drug icariin's induced differentiation of ES-D3 cells Aging curve (see figure 1 ), which is used as the evaluation index of drug efficacy. It can be used for drug development and screening of active ingredients of traditional Chinese medicine. see figure 1 , the effect of icariin on inducing ES cells to differentiate into nerve cells is very obvious, and has a good dose-effect relationship. When it is co-incubated with ES cell embryoid bodies for 8-10 days, more than...
Embodiment 3
[0028] Example 3: The use of icariin in inducing ES cells to differentiate into nerve cells in other aspects
[0029] Using icariin to induce directed differentiation of ES cells into nerve cells, if it can be successfully introduced into the damaged nervous system and work together with host cells, this pathway can become a good way for transplantation to treat neurodegenerative diseases and repair and rebuild damaged nerve tissue. An important source of tissue cells, a large number of nerve cell clones can be cultivated by this method, providing tissue engineering and other research.
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