POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO
DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS

a 2017

POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS

ŠIMARA, Pavel, Irena KRONTORÁD KOUTNÁ, Lenka TESAŘOVÁ and Šimon FARKAŠ

Basic information

Original name

POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS

Authors

ŠIMARA, Pavel, Irena KRONTORÁD KOUTNÁ, Lenka TESAŘOVÁ and Šimon FARKAŠ

Edition

ISSCR 2017 Annual Meeting, 2017

Other information

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10601 Cell biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Poster abstract book ISSCR 2017

Organization unit

Faculty of Informatics

Keywords (in Czech)

hiPSC - endotelialní diferenciace

Keywords in English

hiPSC - endothelial differentiation

Abstract

V originále

Human induced pluripotent stem cells (hiPSCs) are currently considered as the cells which will be widely used in the regeneration of many tissues and organs. One possible utilization is their differentiation into the vascular endothelium. In our laboratory, we have derived hiPSCs from four different cell types. For derivation of hiPSCs we have used as a source cells: neonatal fibroblast, PBMC fraction, HUVECs and HSVECc. On these four hIPSCs lines we tested and evaluated two types of differentiation protocols into endothelial cells. First protocol “A” was aimed on direct differentiation into the lateral mesoderm with subsequent differentiation into the endothelium. In second protocol “B” we have incorporated step of controlled support of differentiation into primitive streak. In case of the first protocol, only up to 31% of cells had desired surface markers (SM) prior to separation. The second protocol yielded up to 48% of cells with desired SM. Quality of differentiated cells in following passages also varied greatly. There was a significant emergence of fibroblast-like cell (FLC) in cell culture by protocol “A”. In such cases, if endothelial cells are not rescued via separation based on SM CD31/CD144 they are overgrown by FLC in matter of days. In passages following initial separation, the cells had 60-80 % of desired SM for the most part, they also needed to be re-separated to maintain these levels of SM. Even though FLC sometimes appeared in cell cultures by protocol “B”, they grew at much slower pace and the rescue separation was as a result much less urgent. Levels of desired SM were also steadily rising even without separation up to 99%. These cells were positive for SM CD31, CD144, CD304 and CD34. Thus differentiated cells showed 100% efficiency on absorption of LDL and tube formation assay. Among the individual hiPSCs lines, we have not found a fundamental difference in the yield between these two protocols.

Links

GBP302/12/G157, research and development project

Name: Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

Investor: Czech Science Foundation

NV16-31501A, research and development project

Name: Tkáňové inženýrství epitelů: Buňky a protokoly pro regenerativní medicínu

Citovat

ŠIMARA, Pavel, Irena KRONTORÁD KOUTNÁ, Lenka TESAŘOVÁ and Šimon FARKAŠ. POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS. In ISSCR 2017 Annual Meeting. 2017.

@proceedings{1384176,
   author = {Šimara, Pavel and Krontorád Koutná, Irena and Tesařová, Lenka and Farkaš, Šimon},
   booktitle = {ISSCR 2017 Annual Meeting},
   keywords = {hiPSC - endothelial differentiation},
   language = {eng},
   title = {POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS},
   url = {http://www.isscr.org/docs/default-source/am17/64553-isscr-abstracts-proof-3-w-cover.pdf?sfvrsn=2},
   year = {2017}
}

TY  - CONF
ID  - 1384176
AU  - Šimara, Pavel - Krontorád Koutná, Irena - Tesařová, Lenka - Farkaš, Šimon
PY  - 2017
TI  - POTENTIAL OF hiPSCs DERIVED FROM DIFFERENT CELL TYPES TO DIFFERENTIATE INTO FUNCTIONAL ENDOTHELIAL CELLS
KW  - hiPSC - endothelial differentiation
UR  - http://www.isscr.org/docs/default-source/am17/64553-isscr-abstracts-proof-3-w-cover.pdf?sfvrsn=2
N2  - Human induced pluripotent stem cells (hiPSCs) are currently considered as the cells which will be widely used in the regeneration of many tissues and organs. One possible utilization is their differentiation into the vascular endothelium. In our laboratory, we have derived hiPSCs from four different cell types. For derivation of hiPSCs we have used as a source cells: neonatal fibroblast, PBMC fraction, HUVECs and HSVECc. On these four hIPSCs lines we tested and evaluated two types of differentiation protocols into endothelial cells. First protocol “A” was aimed on direct differentiation into the lateral mesoderm with subsequent differentiation into the endothelium. In second protocol “B” we have incorporated step of controlled support of differentiation into primitive streak. In case of the first protocol, only up to 31% of cells had desired surface markers (SM) prior to separation. The second protocol yielded up to 48% of cells with desired SM. Quality of differentiated cells in following passages also varied greatly. There was a significant emergence of fibroblast-like cell (FLC) in cell culture by protocol “A”. In such cases, if endothelial cells are not rescued via separation based on SM CD31/CD144 they are overgrown by FLC in matter of days. In passages following initial separation, the cells had 60-80 % of desired SM for the most part, they also needed to be re-separated to maintain these levels of SM. Even though FLC sometimes appeared in cell cultures by protocol “B”, they grew at much slower pace and the rescue separation was as a result much less urgent. Levels of desired SM were also steadily rising even without separation up to 99%. These cells were positive for SM CD31, CD144, CD304 and CD34. Thus differentiated cells showed 100% efficiency on absorption of LDL and tube formation assay. Among the individual hiPSCs lines, we have not found a fundamental difference in the yield between these two protocols.
ER  -

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