Study on the effects of conditioned media from Endothelial Progenitor Cells on rat brain microvascular endothelial cells
Study on the effects of conditioned media from Endothelial Progenitor Cells on rat brain microvascular endothelial cells
A bachelor semester report submitted to the
Lausitz University of Applied Sciences
Written by
Date of submission:
15th of January, 2013
ii
Table of Content
1. Introduction ................................................................................................................................................9
1.1. Endothelial cells
9
1.2. Rat Brain capillary endothelial cells
10
1.3. Groups of Endothelial Progenitor Cells- Conditioned Media
11
1.3.a. Conditioned media
1.3.b. Endothelial Progenitor cells
1.4.
Characterization of EPC
1.4.a. Von Willebrand factor
1.4.b. Fluorescein isothiocyanate (FITC) labelled lectins
1.5.
1.5.a.
1.5.b.
1.5.c.
1.5.d.
12
12
13
14
14
Recent studies employing stem- and progenitor cells
14
Role of endothelial progenitor cells during tissue repair
Effects of EPC on Atherosclerosis
Conditioned media of EPC as a new therapeutic strategy
Paracrine factors
14
16
16
16
2. Aim ............................................................................................................................................................ 20
3. Material and methods .............................................................................................................................. 21
3.1. Suppliers and compositions
21
3.1.a. Chemicals, Kits and basic commodities
3.1.b. Buffers and media
3.1.c. Cells
3.2.
3.3.
3.4.
3.4.a.
3.4.b.
3.4.c.
3.4.d.
3.4.e.
3.5.
Schematic experimental set up
Cultivation of rBCEC-4
Isolation of Mononuclear cells and consequential preparation of CM
Coating of plates for EPC culture
Mononuclear cell isolation from human peripheral/ umbilical cord blood
EPC- medium changes for CM incubation
Collection of EPC-CM
Immunohistochemical stainings of EPC
Proliferation Assays
3.5.a. Presto Blue
A bachelor semester report submitted to the
Lausitz University of Applied Sciences
Written by
Date of submission:
15th of January, 2013
ii
Table of Content
1. Introduction ................................................................................................................................................9
1.1. Endothelial cells
9
1.2. Rat Brain capillary endothelial cells
10
1.3. Groups of Endothelial Progenitor Cells- Conditioned Media
11
1.3.a. Conditioned media
1.3.b. Endothelial Progenitor cells
1.4.
Characterization of EPC
1.4.a. Von Willebrand factor
1.4.b. Fluorescein isothiocyanate (FITC) labelled lectins
1.5.
1.5.a.
1.5.b.
1.5.c.
1.5.d.
12
12
13
14
14
Recent studies employing stem- and progenitor cells
14
Role of endothelial progenitor cells during tissue repair
Effects of EPC on Atherosclerosis
Conditioned media of EPC as a new therapeutic strategy
Paracrine factors
14
16
16
16
2. Aim ............................................................................................................................................................ 20
3. Material and methods .............................................................................................................................. 21
3.1. Suppliers and compositions
21
3.1.a. Chemicals, Kits and basic commodities
3.1.b. Buffers and media
3.1.c. Cells
3.2.
3.3.
3.4.
3.4.a.
3.4.b.
3.4.c.
3.4.d.
3.4.e.
3.5.
Schematic experimental set up
Cultivation of rBCEC-4
Isolation of Mononuclear cells and consequential preparation of CM
Coating of plates for EPC culture
Mononuclear cell isolation from human peripheral/ umbilical cord blood
EPC- medium changes for CM incubation
Collection of EPC-CM
Immunohistochemical stainings of EPC
Proliferation Assays
3.5.a. Presto Blue