Comparison of different methods for the isolation of mesenchymal stem cells from umbilical cord matrix: Proliferation and multilineage differentiation as compared to mesenchymal stem cells from umbilical cord blood and bone marrow

Abstract

We have identified the most appropriate method of isolating human umbilical cord matrix-derived mesenchymal stem cells (UCM-MSCs) and compared morphological, phenotypic, proliferative, and differentiation characteristics of UCM-MSCs with bone marrow-derived MSCs (BM-MSCs) and umbilical cord blood-derived MSCs (UCB-MSCs). Three explant culture methods and 3 enzymatic methods were compared with regards to time for primary culture, cell number, cell morphology, immune phenotype, and differentiation potential. Morphological, phenotypic, proliferative, and differentiation characteristics of UCM-MSCs, BM-MSCs, and UCB-MSCs were also compared. UCM-MSCs isolated using the 10 mm size tissue explant method led to shorter primary culture time, higher numbers of isolated cells, and higher proliferation rates compared with other isolation methods. Immune phenotype and multilineage differentiation capacity did not differ significantly among 6 groups. UCM-MSCs had similar characteristics as BM-MSCs and UCB-MSCs, including fibroblastic morphology, typical immunophenotypic markers, and multilineage differentiation capacity. In comparison with UCB-MSCs and BM-MSCs, UCM-MSCs have higher proliferative capacity, higher rate of chondrogenic differentiation, and higher expression of CD 146. The results suggest that the 10 mm size tissue culture method is the optimal protocol for the isolation of UCM-MSCs. Given the distinct advantages of UC, such as accessibility, painless acquisition, and abundance of cells obtained, we propose that UC be considered an alternative to BM and UCB as a source of MSCs for cell therapy.

Mesenchymal Stem Cells Improve Healing of Diabetic Foot Ulcer

“…Thus, BM-MSCs transplantation is a safe way for DFU, and intramuscular transplantation has been proved to have the best efficacy [22]. However, the number and differentiated potential of BM-MSCs decline with aging [52]. …”

“…UCB-MSCs have a similar morphology, cell surface antigens, and the potential of differentiation into BM-MSCs and umbilical cord-derived mesenchymal stem cells (UC-MSCs) [42, 52, 68]. Additionally, UCB-MSCs have several advantages, such as short doubling time [69], long viable time, and anti-inflammatory activity [42].…”

“…UC-MSCs are generally considered to be rich, safe, of short doubling time, and easy to collect [52]. Compared to BM-MSCs, it has been well documented that UC-MSCs have similar characteristics involving fibroblastic morphology, typical immunophenotypic markers, and multiple differentiation potential to BM-MSCs [79–82].…”

Isolation and characterization of human mesenchymal stem cells derived from synovial fluid by magnetic‐activated cell sorting (MACS)

“…The primary obstacle associated with autologous chondrocyte implantation in articular cartilage repair is the loss of the chondrocyte phenotype (Duan et al, 2015). Prior work has demonstrated that MSCs can be isolated from various types of adult mesenchymal tissue, including bone marrow, skeletal muscle, adipose tissue, and human umbilical cord (Campagnoli et al, 2001;Zuk et al, 2002;Da Silva Meirelles et al, 2006;Kestendjieva et al, 2008;Hua et al, 2013). This is largely due to their high expansion efficiency and their capacity to differentiate into multiple cell types, including adipocytes, osteoblasts, chondrocytes, skeletal myocytes, and tenocytes (Pittenger et al, 1999).…”

Novel clinical uses for cord blood derived mesenchymal stromal cells

“…However, the frequency of MSCs is low, and only a third of CB cells provide a source for generation of MSCs in vitro. Alternative approaches for isolation of MSCs have been developed from the CT [24]. In fact some CB banks now routinely store both CB and CT, offering a unique and potentially advantageous source of HSCs, HPCs and MSCs from a single matched source.…”