Human Platelet Lysate Maintains Stemness of Umbilical Cord-Derived Mesenchymal Stromal Cells and Promote Lung Repair in Rat Bronchopulmonary Dysplasia.

Guilian Liao,Junyuan Hu,Danling Cheng,Duanduan Li,Yan Liao,Guifang Zeng,Muyun Liu,Zan Tang,Xiao Liang,Shiduo Wu,Qiuxing Ouyang,Shaokun Xu,Zeqin Fu

doi:10.3389/fcell.2021.722953

Abstract

Mesenchymal stromal cells (MSCs) show potential for treating preclinical models of newborn bronchopulmonary dysplasia (BPD), but studies of their therapeutic effectiveness have had mixed results, in part due to the use of different media supplements for MSCs expansion in vitro. The current study sought to identify an optimal culture supplement of umbilical cord-derived MSCs (UC-MSCs) for BPD therapy. In this study, we found that UC-MSCs cultured with human platelet lysate (hPL-UCMSCs) were maintained a small size from Passage 1 (P1) to P10, while UC-MSCs cultured with fetal bovine serum (FBS-UCMSCs) became wide and flat. Furthermore, hPL was associated with lower levels of senescence in UC-MSCs during in vitro expansion compared with FBS, as indicated by the results of β-galactosidase staining and measures of senescence-related genes (CDKN2A, CDKN1A, and mTOR). In addition, hPL enhanced the proliferation and cell viability of the UC-MSCs and reduced their doubling time in vitro. Compared with FBS-UCMSCs, hPL-UCMSCs have a greater potential to differentiate into osteocytes and chondrocytes. Moreover, using hPL resulted in greater expression of Nestin and specific paracrine factors (VEGF, TGF-β1, FGF2, IL-8, and IL-6) in UC-MSCs compared to using FBS. Critically, we also found that hPL-UCMSCs are more effective than FBS-UCMSCs for the treatment of BPD in a rat model, with hPL leading to improvements in survival rate, lung architecture and fibrosis, and lung capillary density. Finally, qPCR of rat lung mRNA demonstrated that hPL-UCMSCs had lower expression levels of inflammatory factors (TNF-α and IL-1β) and a key chemokine (MCP-1) at postnatal day 10, and there was significant reduction of CD68+ macrophages in lung tissue after hPL-UCMSCs transplantation. Altogether, our findings suggest that hPL is an optimal culture supplement for UC-MSCs expansion in vitro, and that hPL-UCMSCs promote lung repair in rat BPD disease.

Highlights

Bronchopulmonary dysplasia (BPD) is a serious and common complication of prematurity (Jobe and Bancalari, 2001; Kinsella et al, 2006), which requires mechanical ventilation and oxygen therapy (Baraldi and Filippone, 2007)
We provide evidence that human platelet lysate (hPL) is useful for UC-mesenchymal stem/stromal cells (MSCs) expansion and rejuvenation in vitro, and has superior therapeutic efficacy compared to fetal bovine serum (FBS) in an in vivo rat BPD model
Passage 1 (P1), P4, and P10 MSCs were used for showing their morphology, P10 cells were used for apoptosis and senescence analyses, and P4 cell populations were mainly used for the following in vitro and in vivo experiments

Summary

Introduction

Bronchopulmonary dysplasia (BPD) is a serious and common complication of prematurity (Jobe and Bancalari, 2001; Kinsella et al, 2006), which requires mechanical ventilation and oxygen therapy (Baraldi and Filippone, 2007) It is characterized by a prominent inflammatory response in the lungs that causes restricted lung growth (tissue simplification), subdued alveolar and blood vessel development, and dramatically impaired pulmonary function (Khemani et al, 2007; del Cerro et al, 2014). A growing number of studies in pulmonary medicine have demonstrated that MSC therapy can ameliorate bleomycin, endotoxin, lipopolysaccharide (LPS), or hyperoxia-induced lung injury through the paracrine pathway These studies demonstrate that MSCs can be used to repair damaged tissue and deliver protection via the secretion of specific growth and immunoprotective factors (Ortiz et al, 2003; Gupta et al, 2007; Mei et al, 2007; Xu et al, 2007; Willis et al, 2018)

Objectives

Methods

Results

Conclusion