STUDIES ON THE EFFECT OF DISINTEGRIN AND/OR MESENCHYMAL STEM CELLS IN MICE LIVER INJURY MODEL
Dalia Abdel-Wahab Mohamed;
Abstract
This study was took place in the research unit of natural toxins, biochemistry department, faculty of medicine, Ain shams university.
The aim of the work is to study the effect of purified disintegrin fraction and / or Mesenchymal Stem Cells on liver regeneration in white mice liver injury model induced by CCl¬4.
The current study includes; purification of Cerastes cersates crude venom according to Zaki et al., (2011) through 3 fractionation steps:
1- Gel filtration chromatography on Sephadex G-100 column using ammonium acetate buffer 0.02 M, pH 4.8. Five fractions were obtained and designated A1, A2, B, C and D. The platelet aggregation inhibitory activity has been detected in fraction A2.
2- Ion exchange chromatography on DEAE Sepharose eluted using a linear gradient of ammonium acetate buffer (0.01 M, pH 6.5 and 0.5 M, pH 4.9).Three peaks were obtained and designated A2a, A2b, and A2c .The platelet aggregation inhibitory activity has been detected in A2c fraction.
3- A2c fraction was auto-proteolysed by incubating it ;45μl (0.75 mg/ml) with 5 μl Tris-HCl (50mM)-CaCl2 (10mM) , pH 8.5 for different time intervals ( 24 hours , 48 hours, 72 hours) at 37°C . Products of Auto-proteolysis were detected and their molecular weights were determined by SDS-disc gel electrophoresis (phosphate buffer 0.2 M, pH 7.2). Fraction A2c incubated at 37 C° for 24 hours,48 hours and 72 hours was auto-proteolysed showing 3 bands of molecular weights ; 19 KDa , 15 KDa and 6 KDa respectively.
4- Re-fractionation of products of auto-proteolysed A2c fraction by Gel filtration chromatography on Sephadex G-50 column using ammonium acetate buffer 0.02 M, pH 4.8 , 3 fractions were obtained and designated P¬1 , P¬2 and P¬3.
The aim of the work is to study the effect of purified disintegrin fraction and / or Mesenchymal Stem Cells on liver regeneration in white mice liver injury model induced by CCl¬4.
The current study includes; purification of Cerastes cersates crude venom according to Zaki et al., (2011) through 3 fractionation steps:
1- Gel filtration chromatography on Sephadex G-100 column using ammonium acetate buffer 0.02 M, pH 4.8. Five fractions were obtained and designated A1, A2, B, C and D. The platelet aggregation inhibitory activity has been detected in fraction A2.
2- Ion exchange chromatography on DEAE Sepharose eluted using a linear gradient of ammonium acetate buffer (0.01 M, pH 6.5 and 0.5 M, pH 4.9).Three peaks were obtained and designated A2a, A2b, and A2c .The platelet aggregation inhibitory activity has been detected in A2c fraction.
3- A2c fraction was auto-proteolysed by incubating it ;45μl (0.75 mg/ml) with 5 μl Tris-HCl (50mM)-CaCl2 (10mM) , pH 8.5 for different time intervals ( 24 hours , 48 hours, 72 hours) at 37°C . Products of Auto-proteolysis were detected and their molecular weights were determined by SDS-disc gel electrophoresis (phosphate buffer 0.2 M, pH 7.2). Fraction A2c incubated at 37 C° for 24 hours,48 hours and 72 hours was auto-proteolysed showing 3 bands of molecular weights ; 19 KDa , 15 KDa and 6 KDa respectively.
4- Re-fractionation of products of auto-proteolysed A2c fraction by Gel filtration chromatography on Sephadex G-50 column using ammonium acetate buffer 0.02 M, pH 4.8 , 3 fractions were obtained and designated P¬1 , P¬2 and P¬3.
Other data
| Title | STUDIES ON THE EFFECT OF DISINTEGRIN AND/OR MESENCHYMAL STEM CELLS IN MICE LIVER INJURY MODEL | Other Titles | دراسة تأثير شبيه الديسنتجرين أو / و الخلايا الجذعية على الكبد التالف لفئران التجارب | Authors | Dalia Abdel-Wahab Mohamed | Issue Date | 2014 |
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