Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers?
Human keratinocyte culture has provided the means to treat burns, wounds and skin pathologies. To date, to efficiently culture keratinocytes, cells are cultured on an irradiated feeder layer (iFL), either comprising human (iHFL) or murine (i3T3FL) fibroblasts, and the culture medium is supplemented...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2018-07-01
|
Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | http://www.mdpi.com/1422-0067/19/8/2174 |
id |
doaj-8275b012dd2942d6a034202f792499c9 |
---|---|
record_format |
Article |
spelling |
doaj-8275b012dd2942d6a034202f792499c92020-11-24T23:29:37ZengMDPI AGInternational Journal of Molecular Sciences1422-00672018-07-01198217410.3390/ijms19082174ijms19082174Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers?Sergio Cortez Ghio0Laurence Cantin-Warren1Rina Guignard2Danielle Larouche3Lucie Germain4Université Laval Research Center on Experimental Organogenesis/LOEX, Québec, QC G1J 1Z4, CanadaUniversité Laval Research Center on Experimental Organogenesis/LOEX, Québec, QC G1J 1Z4, CanadaUniversité Laval Research Center on Experimental Organogenesis/LOEX, Québec, QC G1J 1Z4, CanadaUniversité Laval Research Center on Experimental Organogenesis/LOEX, Québec, QC G1J 1Z4, CanadaUniversité Laval Research Center on Experimental Organogenesis/LOEX, Québec, QC G1J 1Z4, CanadaHuman keratinocyte culture has provided the means to treat burns, wounds and skin pathologies. To date, to efficiently culture keratinocytes, cells are cultured on an irradiated feeder layer (iFL), either comprising human (iHFL) or murine (i3T3FL) fibroblasts, and the culture medium is supplemented with a cyclic adenosine monophosphate (cAMP) accumulation inducing agent such as isoproterenol (ISO) or cholera toxin (CT). Previous studies have characterized how the feeder layer type and the cAMP inducer type influence epithelial cells’ phenotype independently from one another, but it is still unknown if an optimal combination of feeder layer and cAMP inducer types exists. We used sophisticated statistical models to search for a synergetic effect of feeder layer and cAMP inducer types on human keratinocytes’ proliferative potential. Our data suggests that, when culturing human keratinocytes, using iHFL over i3T3FL increases population doublings and colony-forming efficiency through signaling pathways involving Ak mouse strain thymoma (Akt, also known as protein kinase B) isoforms 1 to 3, signal transducer and activator of transcription 5 (STAT5), p53, and adenosine monophosphate activated protein kinase α1 (AMPKα1). Both tested cAMP inducers ISO and CT yielded comparable outcomes. However, no significant synergy between feeder layer and cAMP inducer types was detected. We conclude that, to promote human keratinocyte growth in the early passages of culture, co-culturing them with a human feeder layer is preferable to a murine feeder layer.http://www.mdpi.com/1422-0067/19/8/2174keratinocytefeeder layercholera toxinisoproterenolcell culturestatistical model |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sergio Cortez Ghio Laurence Cantin-Warren Rina Guignard Danielle Larouche Lucie Germain |
spellingShingle |
Sergio Cortez Ghio Laurence Cantin-Warren Rina Guignard Danielle Larouche Lucie Germain Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers? International Journal of Molecular Sciences keratinocyte feeder layer cholera toxin isoproterenol cell culture statistical model |
author_facet |
Sergio Cortez Ghio Laurence Cantin-Warren Rina Guignard Danielle Larouche Lucie Germain |
author_sort |
Sergio Cortez Ghio |
title |
Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers? |
title_short |
Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers? |
title_full |
Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers? |
title_fullStr |
Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers? |
title_full_unstemmed |
Are the Effects of the Cholera Toxin and Isoproterenol on Human Keratinocytes’ Proliferative Potential Dependent on Whether They Are Co-Cultured with Human or Murine Fibroblast Feeder Layers? |
title_sort |
are the effects of the cholera toxin and isoproterenol on human keratinocytes’ proliferative potential dependent on whether they are co-cultured with human or murine fibroblast feeder layers? |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2018-07-01 |
description |
Human keratinocyte culture has provided the means to treat burns, wounds and skin pathologies. To date, to efficiently culture keratinocytes, cells are cultured on an irradiated feeder layer (iFL), either comprising human (iHFL) or murine (i3T3FL) fibroblasts, and the culture medium is supplemented with a cyclic adenosine monophosphate (cAMP) accumulation inducing agent such as isoproterenol (ISO) or cholera toxin (CT). Previous studies have characterized how the feeder layer type and the cAMP inducer type influence epithelial cells’ phenotype independently from one another, but it is still unknown if an optimal combination of feeder layer and cAMP inducer types exists. We used sophisticated statistical models to search for a synergetic effect of feeder layer and cAMP inducer types on human keratinocytes’ proliferative potential. Our data suggests that, when culturing human keratinocytes, using iHFL over i3T3FL increases population doublings and colony-forming efficiency through signaling pathways involving Ak mouse strain thymoma (Akt, also known as protein kinase B) isoforms 1 to 3, signal transducer and activator of transcription 5 (STAT5), p53, and adenosine monophosphate activated protein kinase α1 (AMPKα1). Both tested cAMP inducers ISO and CT yielded comparable outcomes. However, no significant synergy between feeder layer and cAMP inducer types was detected. We conclude that, to promote human keratinocyte growth in the early passages of culture, co-culturing them with a human feeder layer is preferable to a murine feeder layer. |
topic |
keratinocyte feeder layer cholera toxin isoproterenol cell culture statistical model |
url |
http://www.mdpi.com/1422-0067/19/8/2174 |
work_keys_str_mv |
AT sergiocortezghio aretheeffectsofthecholeratoxinandisoproterenolonhumankeratinocytesproliferativepotentialdependentonwhethertheyarecoculturedwithhumanormurinefibroblastfeederlayers AT laurencecantinwarren aretheeffectsofthecholeratoxinandisoproterenolonhumankeratinocytesproliferativepotentialdependentonwhethertheyarecoculturedwithhumanormurinefibroblastfeederlayers AT rinaguignard aretheeffectsofthecholeratoxinandisoproterenolonhumankeratinocytesproliferativepotentialdependentonwhethertheyarecoculturedwithhumanormurinefibroblastfeederlayers AT daniellelarouche aretheeffectsofthecholeratoxinandisoproterenolonhumankeratinocytesproliferativepotentialdependentonwhethertheyarecoculturedwithhumanormurinefibroblastfeederlayers AT luciegermain aretheeffectsofthecholeratoxinandisoproterenolonhumankeratinocytesproliferativepotentialdependentonwhethertheyarecoculturedwithhumanormurinefibroblastfeederlayers |
_version_ |
1725544637411622912 |