Venom-based peptide and application thereof
A venom and amino acid technology, applied in the direction of peptides, embryonic cells, germ cells, etc., to achieve the effect of promoting self-renewal
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Embodiment 1
[0092] This example provides venom-based peptide P13 and mutant M6 of P13.
[0093] The amino acid sequence of P13 is shown in SEQ ID No.1, and the amino acid sequence of M6 is shown in SEQ ID No.2.
[0094] P13 is mutated to M6:
[0095] P13 was mutated to synthesize its derivative P13-M5 (sequence shown in SEQ ID No.3). It was reported in the literature that the binding affinity of the basic amino acid arginine to heparin is 2.5 times that of lysine, so further mutations resulted in P13-M6.
Embodiment 2
[0097] This embodiment provides a medium for culturing human embryonic stem cells, specifically as follows.
[0098] (1) E6 and P13;
[0099] (2) E6 and M6;
[0100] (3) E8 and P13;
[0101] (4) E8 and M6.
[0102] It should be noted that in (1) to (4), the concentrations of P13 and M6 were 100 μM. Specifically, the E6 medium includes: DMEM / F12, insulin, L-ascorbic acid, selenium, NaHCO 3 and transferrin;
[0103] E8 medium includes: DMEM / F12, FGF2, TGFβ1, insulin, L-ascorbic acid, selenium, NaHCO 3 and transferrin.
[0104] Human ES cell culture
[0105] Taking E8 medium as an example, human ES cells (hESC, H1) were maintained in E8 medium using matrigel-coated tissue culture plates. E8 medium has eight components: DMEM / F12, L-ascorbic acid-2-magnesium phosphate (64mg / L), sodium selenium (14μg / L), insulin (19.4mg / L), NaHCO3 (543mg / L), Transferrin (10.7 mg / L) and FGF2 (100 μg / L) and TGFβ1 (2 μg / L) growth factors.
[0106] Once the cells reached 80-85% confluency, cel...
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