Abstract

To date, various types of cells for seeding regenerative scaffolds have been used for bone tissue engineering. Among seed cells, the mesenchymal stem cells derived from human umbilical cord Wharton’s jelly (hUCMSCs) represent a promising candidate and hold potential for bone tissue engineering due to the the lack of ethical controversies, accessibility, sourced by non-invasive procedures for donors, a reduced risk of contamination, osteogenic differentiation capacities, and higher immunomodulatory capacity. However, the current culture methods are somewhat complicated and inefficient and often fail to make the best use of the umbilical cord (UC) tissues. Moreover, these culture processes cannot be performed on a large scale and under strict quality control. As a result, only a small quantity of cells can be harvested using the current culture methods. To solve these problems, we designed and evaluated an UC Wharton’s jelly repeated culture device. Using this device, hUCMSCs were obtained from the repeated cultures and their quantities and biological characteristics were compared. We found that using our culture device, which retained all tissue blocks on the bottom of the dish, the total number of obtained cells increased 15–20 times, and the time required for the primary passage was reduced. Moreover, cells harvested from the repeated cultures exhibited no significant difference in their immunophenotype, potential for multilineage differentiation, or proliferative, osteoinductive capacities, and final osteogenesis. The application of the repeated culture frame (RCF) not only made full use of the Wharton’s jelly but also simplified and specified the culture process, and thus, the culture efficiency was significantly improved. In summary, abundant hUCMSCs of dependable quality can be acquired using the RCF.

Highlights

  • Tissue-engineered bones (TEBs) have been widely researched for facilitating bone repair, within a broad range of clinical applications [1,2,3]

  • As proposed by the mesenchymal and tissue stem cell committee of the International Society for Cellular Therapy (ISCT), cells must be positive for CD73, CD90 and CD105 to be defined as mesenchymal stem cells (MSCs) [17]

  • IF staining of freshly excised umbilical cord (UC) showed that the CD73+, CD90+ and CD105+ MSCs were mainly located in the Wharton’s jelly (Fig. 1J–L) and that the CD31+ endothelial progenitor cells were in the vascular intima

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Summary

Introduction

Tissue-engineered bones (TEBs) have been widely researched for facilitating bone repair, within a broad range of clinical applications [1,2,3]. Due to their self-renewal and multipotent capacities, mesenchymal stem cells (MSCs) are considered the ‘gold standard’ seed cells for constructing TEBs. To date, MSCs derived from bone marrow and adipose tissue have been widely applied in bone tissue engineering [4,5]. There are still some drawbacks regarding the applications of bone marrow- or adipose-derived mesenchymal stem cells (BMSCs or ADSCs), including the low number of MSCs in marrow or fat, and the fact that with aging, their proliferative and differentiation capacities decrease significantly [6]. Seeking alternative types of MSCs, with wider availability, an easier harvesting procedure, and more robust proliferative and differentiation abilities has become an urgent issue

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