Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche

Hristina Obradovic; Jelena Krstic; Drenka Trivanovic; Slavko Mojsilovic; Ivana Okic; Tamara Kukolj; Vesna Ilic; Aleksandra Jaukovic; Milan Terzic; Diana Bugarski

doi:10.1016/j.placenta.2019.05.005

Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche

Hristina Obradovic, Jelena Krstic, Drenka Trivanovic, Slavko Mojsilovic, Ivana Okic, Tamara Kukolj, Vesna Ilic, Aleksandra Jaukovic, Milan Terzic, Diana Bugarski

Department of Medicine

Research output: Contribution to journal › Article › peer-review

18 Citations (Scopus)

Abstract

INTRODUCTION: Mesenchymal stem cells from Wharton's Jelly of a human umbilical cord (WJ-MSCs) are a potential tool in regenerative medicine based on their availability, proliferative potential and differentiation capacity. Since their physiological niche contains low oxygen levels, we investigated whether cultivation of WJ-MSCs at 3% O2 affects their main features.

METHODS: WJ-MSCs were cultured under 21% and 3% O2. Proliferation rate was followed by short and long term proliferation assays, clonogenic capacity by CFU-F assay and cell cycle and death by flow cytometry. Differentiation capacity was investigated by histochemical staining after induced differentiation. Pluripotency and differentiation markers' expression was determined by RT-PCR. Migration capacity was followed by scratch assay and mobilization from collagen, and the activity of proteolytic enzymes by zymography. Specific inhibitors of MAPK and Wnt/β-catenin pathways were used to investigate underlying molecular mechanisms.

RESULTS: Compared to standard 21% O2, cultivation of WJ-MSCs at 3% O2 did not influence their immunophenotype, while it modulated their differentiation process and enhanced their clonogenic and expansion capacity. 3% O2 induced transient change in cell cycle and prevented cell death. The expression of NANOG, OCT4A, OCT4B and SOX2 was increased at 3% O2. Both cultivation and preculturing of WJ-MSCs at 3% O2 increased their in vitro migratory capacity and enhanced the activity of proteolytic enzymes. ERK1/2 mediated WJ-MSCs' mobilization from collagen regardless of oxygen levels, while Wnt/β-catenin pathway was activated during migration and mobilization at standard conditions.

CONCLUSION: Culturing of WJ-MSCs under 3% O2 should be considered a credible condition when investigating their properties and potential use.

Original language	English
Pages (from-to)	25-34
Number of pages	10
Journal	Placenta
Volume	82
DOIs	https://doi.org/10.1016/j.placenta.2019.05.005
Publication status	Published - Jul 2019

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Obradovic, H., Krstic, J., Trivanovic, D., Mojsilovic, S., Okic, I., Kukolj, T., Ilic, V., Jaukovic, A., Terzic, M., & Bugarski, D. (2019). Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche. Placenta, 82, 25-34. https://doi.org/10.1016/j.placenta.2019.05.005

Obradovic, H, Krstic, J, Trivanovic, D, Mojsilovic, S, Okic, I, Kukolj, T, Ilic, V, Jaukovic, A, Terzic, M & Bugarski, D 2019, 'Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche', Placenta, vol. 82, pp. 25-34. https://doi.org/10.1016/j.placenta.2019.05.005

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title = "Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche",

abstract = "INTRODUCTION: Mesenchymal stem cells from Wharton's Jelly of a human umbilical cord (WJ-MSCs) are a potential tool in regenerative medicine based on their availability, proliferative potential and differentiation capacity. Since their physiological niche contains low oxygen levels, we investigated whether cultivation of WJ-MSCs at 3% O2 affects their main features.METHODS: WJ-MSCs were cultured under 21% and 3% O2. Proliferation rate was followed by short and long term proliferation assays, clonogenic capacity by CFU-F assay and cell cycle and death by flow cytometry. Differentiation capacity was investigated by histochemical staining after induced differentiation. Pluripotency and differentiation markers' expression was determined by RT-PCR. Migration capacity was followed by scratch assay and mobilization from collagen, and the activity of proteolytic enzymes by zymography. Specific inhibitors of MAPK and Wnt/β-catenin pathways were used to investigate underlying molecular mechanisms.RESULTS: Compared to standard 21% O2, cultivation of WJ-MSCs at 3% O2 did not influence their immunophenotype, while it modulated their differentiation process and enhanced their clonogenic and expansion capacity. 3% O2 induced transient change in cell cycle and prevented cell death. The expression of NANOG, OCT4A, OCT4B and SOX2 was increased at 3% O2. Both cultivation and preculturing of WJ-MSCs at 3% O2 increased their in vitro migratory capacity and enhanced the activity of proteolytic enzymes. ERK1/2 mediated WJ-MSCs' mobilization from collagen regardless of oxygen levels, while Wnt/β-catenin pathway was activated during migration and mobilization at standard conditions.CONCLUSION: Culturing of WJ-MSCs under 3% O2 should be considered a credible condition when investigating their properties and potential use.",

author = "Hristina Obradovic and Jelena Krstic and Drenka Trivanovic and Slavko Mojsilovic and Ivana Okic and Tamara Kukolj and Vesna Ilic and Aleksandra Jaukovic and Milan Terzic and Diana Bugarski",

year = "2019",

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doi = "10.1016/j.placenta.2019.05.005",

language = "English",

volume = "82",

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TY - JOUR

T1 - Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche

AU - Obradovic, Hristina

AU - Krstic, Jelena

AU - Trivanovic, Drenka

AU - Mojsilovic, Slavko

AU - Okic, Ivana

AU - Kukolj, Tamara

AU - Ilic, Vesna

AU - Jaukovic, Aleksandra

AU - Terzic, Milan

AU - Bugarski, Diana

PY - 2019/7

Y1 - 2019/7

N2 - INTRODUCTION: Mesenchymal stem cells from Wharton's Jelly of a human umbilical cord (WJ-MSCs) are a potential tool in regenerative medicine based on their availability, proliferative potential and differentiation capacity. Since their physiological niche contains low oxygen levels, we investigated whether cultivation of WJ-MSCs at 3% O2 affects their main features.METHODS: WJ-MSCs were cultured under 21% and 3% O2. Proliferation rate was followed by short and long term proliferation assays, clonogenic capacity by CFU-F assay and cell cycle and death by flow cytometry. Differentiation capacity was investigated by histochemical staining after induced differentiation. Pluripotency and differentiation markers' expression was determined by RT-PCR. Migration capacity was followed by scratch assay and mobilization from collagen, and the activity of proteolytic enzymes by zymography. Specific inhibitors of MAPK and Wnt/β-catenin pathways were used to investigate underlying molecular mechanisms.RESULTS: Compared to standard 21% O2, cultivation of WJ-MSCs at 3% O2 did not influence their immunophenotype, while it modulated their differentiation process and enhanced their clonogenic and expansion capacity. 3% O2 induced transient change in cell cycle and prevented cell death. The expression of NANOG, OCT4A, OCT4B and SOX2 was increased at 3% O2. Both cultivation and preculturing of WJ-MSCs at 3% O2 increased their in vitro migratory capacity and enhanced the activity of proteolytic enzymes. ERK1/2 mediated WJ-MSCs' mobilization from collagen regardless of oxygen levels, while Wnt/β-catenin pathway was activated during migration and mobilization at standard conditions.CONCLUSION: Culturing of WJ-MSCs under 3% O2 should be considered a credible condition when investigating their properties and potential use.

AB - INTRODUCTION: Mesenchymal stem cells from Wharton's Jelly of a human umbilical cord (WJ-MSCs) are a potential tool in regenerative medicine based on their availability, proliferative potential and differentiation capacity. Since their physiological niche contains low oxygen levels, we investigated whether cultivation of WJ-MSCs at 3% O2 affects their main features.METHODS: WJ-MSCs were cultured under 21% and 3% O2. Proliferation rate was followed by short and long term proliferation assays, clonogenic capacity by CFU-F assay and cell cycle and death by flow cytometry. Differentiation capacity was investigated by histochemical staining after induced differentiation. Pluripotency and differentiation markers' expression was determined by RT-PCR. Migration capacity was followed by scratch assay and mobilization from collagen, and the activity of proteolytic enzymes by zymography. Specific inhibitors of MAPK and Wnt/β-catenin pathways were used to investigate underlying molecular mechanisms.RESULTS: Compared to standard 21% O2, cultivation of WJ-MSCs at 3% O2 did not influence their immunophenotype, while it modulated their differentiation process and enhanced their clonogenic and expansion capacity. 3% O2 induced transient change in cell cycle and prevented cell death. The expression of NANOG, OCT4A, OCT4B and SOX2 was increased at 3% O2. Both cultivation and preculturing of WJ-MSCs at 3% O2 increased their in vitro migratory capacity and enhanced the activity of proteolytic enzymes. ERK1/2 mediated WJ-MSCs' mobilization from collagen regardless of oxygen levels, while Wnt/β-catenin pathway was activated during migration and mobilization at standard conditions.CONCLUSION: Culturing of WJ-MSCs under 3% O2 should be considered a credible condition when investigating their properties and potential use.

U2 - 10.1016/j.placenta.2019.05.005

DO - 10.1016/j.placenta.2019.05.005

M3 - Article

C2 - 31174623

SN - 0143-4004

VL - 82

SP - 25

EP - 34

JO - Placenta

JF - Placenta

ER -

Improving stemness and functional features of mesenchymal stem cells from Wharton's jelly of a human umbilical cord by mimicking the native, low oxygen stem cell niche

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