Comparison of Preterm and Term Wharton’s Jelly-Derived Mesenchymal Stem Cell Properties in Different Oxygen Tensions

Abstract

Mesenchymal stem cells (MSCs) have shown promise as therapeutic agents in treating morbidities associated with premature birth. MSCs derived from the human umbilical cord are easy to isolate and have low immunogenicity and a robust ability to secrete paracrine factors. To date, there are no studies evaluating preterm versus term umbilical cord tissue-derived MSCs. Therefore, our aim was twofold: (1) to compare stem cell properties in preterm versus term MSCs and (2) to examine the impact of oxygen tension on stem cell behavior. Umbilical cord tissue was obtained from 5 preterm and 5 term neonates. The cells were isolated and characterized as MSCs in accordance with the International Society for Cellular Therapy. We exposed MSCs to different oxygen tensions to examine the impact of environmental factors on cell performance. We studied the following stem cell properties: (i) motility, (ii) proliferation, (iii) senescence, (iv) cell viability, (v) colony-forming unit efficiency, and (vi) inflammatory cytokine expression. Under normoxia (21% O₂), cells from preterm and term infants had similar properties. Under hypoxic conditions (1% O₂), term MSCs had better cell proliferation; however, cells exposed to hyperoxia (90% O₂) had the slowest motility and lowest cell viability (p < 0.05). There was no difference in the expression of senescence or cytokine expression between the groups. The term cells demonstrated more colony-forming efficiency than the preterm cells. In sum, our preliminary findings suggest that MSCs derived from term and preterm umbilical cords have similar characteristics, offering the potential of future autologous/allogeneic MSC transplants in neonates.

Intranasal delivery of human umbilical cord Wharton's jelly mesenchymal stromal cells restores lung alveolarization and vascularization in experimental bronchopulmonary dysplasia

“…Cells were negative for hematologic stem cell markers HLA‐DR, CD117, and CD79 alpha. Consistent with the criteria set by International Society for Cell and Gene Therapy, cells differentiated into adipocytes, chondrocytes, and osteocytes when grown with lineage specific media, as previously describe …”

“…Reasons that may explain the effectiveness in our study include: multiple dose administration, less severe BPD model, intranasal volume given, and anatomic positioning of animals during cell delivery . Moreover, using an automated cell counter we tested the viability of the human umbilical cord MSCs just prior to animal administration and our yield ranged between 93% and 99% . Despite being a crude approximation of cell viability, we wanted a quick and objective measure of the percent of live cells we were providing to the animals.…”

“…[40][41][42] Moreover, using an automated cell counter we tested the viability of the human umbilical cord MSCs just prior to animal administration and our yield ranged between 93% and 99%. 20 Despite being a crude approximation of cell viability, we wanted a quick and objective measure of the percent of live cells we were providing to F I G U R E 6 Protein analysis of lung homogenates from room air (RA) control, oxygen injured (bronchopulmonary dysplasia [BPD]), and BPD treated with mesenchymal stromal cells (BPD + MSCs). A, Select data displayed in the following grouping: Pro-inflammatory; Antiinflammatory; Cell survival; Wound healing; Fibrogenesis; and Anti-vascular.…”

“…After tissue enzymatic digestion, cells were expanded on culture dishes under standard conditions. Wharton's jelly cells met the minimum criteria for MSCs, as described in our previous publication …”

“…Wharton's jelly cells met the minimum criteria for MSCs, as described in our previous publication. 20 Preparation of MSCs was achieved as follows, 5 × 10 5 cells were seeded onto 100 mm tissue culture dishes and grown under standard conditions with media changes every 2-3 days. At 24 hours prior to administration, cells were checked for confluency (80%-90%) and the media was changed for the last time.…”