Pressure Stimuli Improve the Proliferation of Wharton's Jelly-Derived Mesenchymal Stem Cells under Hypoxic Culture Conditions.

Sang Eon Park,Hyeongseop Kim,Gyu Ha Ryu,Soojin Kwon,Soo-Young Oh,Jong Wook Chang,Hong Bae Jeon,Suk-Joo Choi

doi:10.3390/ijms21197092

Abstract

Mesenchymal stem cells (MSCs) are safe, and they have good therapeutic efficacy through their paracrine action. However, long-term culture to produce sufficient MSCs for clinical use can result in side-effects, such as an inevitable senescence and the reduction of the therapeutic efficacy of the MSCs. In order to overcome this, the primary culture conditions of the MSCs can be modified to simulate the stem cells’ niche environment, resulting in accelerated proliferation, the achievement of the target production yield at earlier passages, and the improvement of the therapeutic efficacy. We exposed Wharton’s jelly-derived MSCs (WJ-MSCs) to pressure stimuli during the primary culture step. In order to evaluate the proliferation, stemness, and therapeutic efficacy of WJ-MSCs, image, genetic, and Western blot analyses were carried out. Compared with standard incubation culture conditions, the cell proliferation was significantly improved when the WJ-MSCs were exposed to pressure stimuli. However, the therapeutic efficacy (the promotion of cell proliferation and anti-apoptotic effects) and the stemness of the WJ-MSCs was maintained, regardless of the culture conditions. Exposure to pressure stimuli is a simple and efficient way to improve WJ-MSC proliferation without causing changes in stemness and therapeutic efficacy. In this way, clinical-grade WJ-MSCs can be produced rapidly and used for therapeutic applications.

Highlights

Mesenchymal stem cells (MSCs) were first reported in 1960 [1]
In order to assess the proliferation and yield of the Wharton’s jelly-derived MSCs (WJ-MSCs) primary culture, according to incubation conditions, cell confluency was observed by optical microscopy on the 4th, 7th, and 9th days; the cells were collected on the 9th day in order to calculate yields (Figure 1)
When the WJ-MSCs at passage 0 were exposed to standard incubation conditions (5% CO2, 37 ◦C incubator, as a control), or to the 5% O2 hypoxia conditions (C+H), or the 5% O2 hypoxia with 2.0 PSI pressure conditions (C+H+P), confluency changes were observed (Figure 1A)

Summary

Introduction

Mesenchymal stem cells (MSCs) were first reported in 1960 [1]. These cells attach themselves to plastic culture flasks in a similar form to fibroblasts, and are defined as adult stem cells with adipogenic, osteogenic, and chondrogenic properties. MSCs are considered to be a promising treatment for rare and intractable diseases, as these cells have the immune privilege (they do not cause immune reactions when transplanted into a person), and have demonstrated therapeutic efficacy in various diseases through their paracrine action [3,4]. Such secretions of proteins—which maintain patient homeostasis, remove causative substances, and regenerate damaged tissue—are the main therapeutic mechanism of MSCs [5,6]. The dosage of MSCs needs to be relatively high compared to other stem cell therapies

Methods

Results

Conclusion