Generation of Kidney Directed Differentiation Platform from Human Pluripotent Stem Cells / 中国生物化学与分子生物学报

Kidney is an essential organ in human body with multiple physiological functions. However, there is 10 % population worldwide with renal disease. It is urgent to generate a model which is more similar with kidney at structural and functional level to study renal disease. The rise of in vitro differentiation technology from pluripotent stem cells gives regeneration medicine and precise medicine new energy. This study mimics kidney development in vitro by inducing human pluripotent stem cells including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) into kidney progenitor cells, and further forming nephrons, which is the structure and function unit in kidney. Human pluripotent stem cells were differentiated into primitive streak through activating WNT pathway while inhibiting TGF-(B signaling. Afterward, the primitive streak spontaneously differentiated into intermediate mesoderm. Then, we induced intermediate mesoderm cells into kidney progenitor cells through FGF pathway. The FACS analysis data indicated kidney progenitor cells were up to 51. 5%-61. 9% in total cell population. Immuno-stai-ning results showed these structures contained podocytes of glomerulus, proximal tubule, and distal tubule. This kidney differentiation protocol is stable, high-efficient, and well repeatable. This research provides a novel platform for early human kidney development study, kidney disease modeling, and drug screening.

N'-(3-Bromo-4-methoxy-benzyl-idene)nicotinohydrazide monohydrate.

In the title compound, C(14)H(12)BrN(3)O(2)·H(2)O, the benzene ring is oriented at a dihedral angle of 39.66 (11)° with respect to the pyridine ring. The solvent water mol-ecule links with the organic compound via O-H⋯O, O-H⋯N and N-H⋯O hydrogen bonding.

(E)-N'-(1,3-Benzodioxol-5-ylmethyl-ene)nicotinohydrazide monohydrate.

In the title compound, C(14)H(11)N(3)O(3)·H(2)O, the planar [maximum deviation 0.135 (1) Å] 1,3-benzodioxole ring system is oriented at a dihedral angle of 13.93 (7)° with respect to the pyridine ring. Extensive inter-molecular N-H⋯O, O-H⋯O, O-H⋯N and weak C-H⋯O hydrogen bonding is present in the crystal structure.

N'-(Propan-2-yl-idene)nicotinohydrazide.

Crystals of the title compound, C(9)H(11)N(3)O, were obtained from a condensation reaction of nicotinohydrazide and acetone. In the mol-ecular structure, the pyridine ring is oriented at a dihedral angle of 36.28 (10)° with respect to the amide plane. In the crystal structure, mol-ecules are linked via N-H⋯O hydrogen bonds, forming chains.

(E)-N'-(3,4-Dichloro-benzyl-idene)nicotino-hydrazide monohydrate.

In the title compound, C(13)H(9)Cl(2)N(3)O·H(2)O, the 3,4-dichloro-benzene ring is nearly coplanar with the pyridine ring, making a dihedral angle of 4.78 (8)°. Inter-molecular O-H⋯O, O-H⋯N, N-H⋯O and weak C-H⋯O hydrogen bonding is present in the crystal structure.

N'-(3-Phenyl-allyl-idene)isonicotino-hydrazide.

The asymmetric unit of the title compound, C(15)H(13)N(3)O, contains two similar mol-ecules. Each mol-ecule is non-planar, as indicated by the dihedral angles between the pyridine and benzene rings of 45.2 (2) and 56.6 (2)°. The crystal structure is consolidated by inter-molecular N-H⋯O hydrogen bonds.

2,4-Dichloro-benzaldehyde 2,4-dinitro-phenyl-hydrazone.

The asymmetric unit of the title compound, C(13)H(8)Cl(2)N(4)O(4), contains two independent but similar and almost planar mol-ecules. An intra-molecular N-H⋯O hydrogen bond is observed in each mol-ecule.

1-(4-Nitro-phen-yl)-2-(3-phenyl-allyl-idene)hydrazine.

In the title compound, C(15)H(13)N(3)O(2), the nitro-benzene and benzene rings make a dihedral angle of 9.1 (2)°. The crystal structure is consolidated by inter-molecular N-H⋯O hydrogen bonds.

2,6-Dichloro-benzaldehyde oxime.

In the title compound, C(7)H(5)Cl(2)NO, there are two mol-ecules in the asymmetric unit. The mol-ecules are essentially identical. Each mol-ecule is connected to a symmetry-related mol-ecule through an inversion center by O-H⋯N hydrogen bonds, building an R(2) (2)(6) graph-set motif.

Expression, Purification and Characterization of Gly14-Humanin, a Novel Neuroprotective Peptide / 中国生物工程杂志

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Expression, Purification and Characterization of Gly14-Humanin, a Novel Neuroprotective Peptide / 中国生物工程杂志

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Expression,Purification and Characterization of [Gly14]-Humanin,a Novel Neuroprotective Peptide / 中国生物工程杂志

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.