A novel method for the collection of nanoscopic vesicles from an organotypic culture model

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A novel method for the collection of nanoscopic vesicles from an organotypic culture model. / Iordachescu, Alexandra; Hulley, Philippa; Grover, Liam m.

In: RSC Advances, Vol. 8, No. 14, 2018, p. 7622-7632.

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Iordachescu, Alexandra ; Hulley, Philippa ; Grover, Liam m. / A novel method for the collection of nanoscopic vesicles from an organotypic culture model. In: RSC Advances. 2018 ; Vol. 8, No. 14. pp. 7622-7632.

Bibtex

@article{c7babb6959634da892490b3396649cf6,
title = "A novel method for the collection of nanoscopic vesicles from an organotypic culture model",
abstract = "Nanovesicles, exosomes and other membrane bound particles excreted by cells are currently gaining research attention since they have been shown to play a significant role in many biologically related processes. Vesicles are now thought to mediate cellular communication, transmission of some diseases and pathologically mediated calcification. Matrix vesicles have long been proposed to be central to the controlled mineralisation of bone. They remain relatively poorly studied, however, since they are challenging to extract from biological media. One difficulty is the presence of a mineral content in comparison to pure lipid vesicles, meaning that standard separation process such as ultracentrifugation are unable to precisely separate on the basis of size or weight. In this paper we report the separation of matrix vesicles from an organotypic bone culture system using a process of immunoprecipitation. Matrix vesicles were extracted using polymeric beads that were modified with an antibody for tissue non-specific alkaline phosphatase (TNALP), a surface marker abundant in bone-derived vesicles. The vesicles isolated were positive for adenosine triphosphate (ATP), the substrate for TNALP and were demonstrated to have a high-binding affinity to type I collagen, the principal collagen type found in bone. This protocol enables more detailed study of the process and regulation of mineralisation.",
author = "Alexandra Iordachescu and Philippa Hulley and Liam m. Grover",
year = "2018",
doi = "10.1039/C7RA12511A",
language = "English",
volume = "8",
pages = "7622--7632",
journal = "RSC Advances",
issn = "2046-2069",
publisher = "Royal Society of Chemistry",
number = "14",

}

RIS

TY - JOUR

T1 - A novel method for the collection of nanoscopic vesicles from an organotypic culture model

AU - Iordachescu, Alexandra

AU - Hulley, Philippa

AU - Grover, Liam m.

PY - 2018

Y1 - 2018

N2 - Nanovesicles, exosomes and other membrane bound particles excreted by cells are currently gaining research attention since they have been shown to play a significant role in many biologically related processes. Vesicles are now thought to mediate cellular communication, transmission of some diseases and pathologically mediated calcification. Matrix vesicles have long been proposed to be central to the controlled mineralisation of bone. They remain relatively poorly studied, however, since they are challenging to extract from biological media. One difficulty is the presence of a mineral content in comparison to pure lipid vesicles, meaning that standard separation process such as ultracentrifugation are unable to precisely separate on the basis of size or weight. In this paper we report the separation of matrix vesicles from an organotypic bone culture system using a process of immunoprecipitation. Matrix vesicles were extracted using polymeric beads that were modified with an antibody for tissue non-specific alkaline phosphatase (TNALP), a surface marker abundant in bone-derived vesicles. The vesicles isolated were positive for adenosine triphosphate (ATP), the substrate for TNALP and were demonstrated to have a high-binding affinity to type I collagen, the principal collagen type found in bone. This protocol enables more detailed study of the process and regulation of mineralisation.

AB - Nanovesicles, exosomes and other membrane bound particles excreted by cells are currently gaining research attention since they have been shown to play a significant role in many biologically related processes. Vesicles are now thought to mediate cellular communication, transmission of some diseases and pathologically mediated calcification. Matrix vesicles have long been proposed to be central to the controlled mineralisation of bone. They remain relatively poorly studied, however, since they are challenging to extract from biological media. One difficulty is the presence of a mineral content in comparison to pure lipid vesicles, meaning that standard separation process such as ultracentrifugation are unable to precisely separate on the basis of size or weight. In this paper we report the separation of matrix vesicles from an organotypic bone culture system using a process of immunoprecipitation. Matrix vesicles were extracted using polymeric beads that were modified with an antibody for tissue non-specific alkaline phosphatase (TNALP), a surface marker abundant in bone-derived vesicles. The vesicles isolated were positive for adenosine triphosphate (ATP), the substrate for TNALP and were demonstrated to have a high-binding affinity to type I collagen, the principal collagen type found in bone. This protocol enables more detailed study of the process and regulation of mineralisation.

U2 - 10.1039/C7RA12511A

DO - 10.1039/C7RA12511A

M3 - Article

VL - 8

SP - 7622

EP - 7632

JO - RSC Advances

JF - RSC Advances

SN - 2046-2069

IS - 14

ER -