Enzymatic Isolation, Amplification and Characterization of Dental Pulp Stem Cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00179906%3A_____%2F19%3A10398958" target="_blank" >RIV/00179906:_____/19:10398958 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11150/19:10398958

Výsledek na webu

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=5iJoHjodRI" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=5iJoHjodRI</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Enzymatic Isolation, Amplification and Characterization of Dental Pulp Stem Cells

Popis výsledku v původním jazyce

The dental pulp represents an easily acces-sible source of adult dental pulp stem cells (DPSCs). The preferred approach to DPSC isolation is enzy-matic digestion. However, the duration of the enzymat-ic activity is crucial. The purpose of this study was to isolate the DPSC populations using this method, characterize their biological properties and prolifer-ation capacity, and to determine their ability to dif-ferentiate into mature cells. Before enzymatic diges-tion using 0.05% trypsin, we used the homogenization method in order to obtain a fine homogenate from the solid pulp tissue. The stem cells were cultivated in modified cultivation medium for mesenchymal adult progenitor cells containing 2% foetal bovine serum, growth factors and insulin-transferrin-sele-nium supplement. We were successfully able to iso-late 10 populations of DPSCs. The vitality of DPSCs did not drop below 90 %. However, the DPSCs showed a significant decrease in the relative telomere length number with increasing passaging (P &lt; 0.05). Isolated DPSCs highly expressed the CD markers: CD29, CD44, CD90, CD13, CD73 and CD166. In con-trast, CD markers CD31, CD106, CD34 and CD45 were negative or low positive. We confirmed the high osteogenic and chondrogenic potential of the isolated stem cells. Isolated DPSCs did not show signs of cell degeneration or spontaneous differentiation during the entire cultivation. In addition, we were able to shorten the enzyme activity duration, and we were the first to demonstrate trypsin as the enzyme used for the enzymatic digestion method with the viability over 90 % of isolated DPSCs using this method.

Název v anglickém jazyce

Enzymatic Isolation, Amplification and Characterization of Dental Pulp Stem Cells

Popis výsledku anglicky

The dental pulp represents an easily acces-sible source of adult dental pulp stem cells (DPSCs). The preferred approach to DPSC isolation is enzy-matic digestion. However, the duration of the enzymat-ic activity is crucial. The purpose of this study was to isolate the DPSC populations using this method, characterize their biological properties and prolifer-ation capacity, and to determine their ability to dif-ferentiate into mature cells. Before enzymatic diges-tion using 0.05% trypsin, we used the homogenization method in order to obtain a fine homogenate from the solid pulp tissue. The stem cells were cultivated in modified cultivation medium for mesenchymal adult progenitor cells containing 2% foetal bovine serum, growth factors and insulin-transferrin-sele-nium supplement. We were successfully able to iso-late 10 populations of DPSCs. The vitality of DPSCs did not drop below 90 %. However, the DPSCs showed a significant decrease in the relative telomere length number with increasing passaging (P &lt; 0.05). Isolated DPSCs highly expressed the CD markers: CD29, CD44, CD90, CD13, CD73 and CD166. In con-trast, CD markers CD31, CD106, CD34 and CD45 were negative or low positive. We confirmed the high osteogenic and chondrogenic potential of the isolated stem cells. Isolated DPSCs did not show signs of cell degeneration or spontaneous differentiation during the entire cultivation. In addition, we were able to shorten the enzyme activity duration, and we were the first to demonstrate trypsin as the enzyme used for the enzymatic digestion method with the viability over 90 % of isolated DPSCs using this method.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

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OECD FORD obor

10601 - Cell biology

Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Folia Biologica

ISSN

0015-5500

e-ISSN

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Svazek periodika

65

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

10

Strana od-do

124-133

Kód UT WoS článku

000500271800002

EID výsledku v databázi Scopus

2-s2.0-85073656780