Isolation of Mesenchymal Stem Cells from Cryopreserved Human Umbilical Cord Blood

Abstract

Umbilical cord blood (UCB) is well known to be a rich source of hematopoietic stem cells with practical and ethical advantages. Because mesenchymal stem cells (MSCs) from bone marrow have been regarded as good materials for cell/gene therapy as well as for tissue engineering because of their multidifferentiation potential, a number of trials have been undertaken to isolate MSCs from UCB. However, the results have been controversial, and little has become known about the effect of cryopreservation on the isolation of these stem cells. In this study, we examined the ability of cryopreserved UCB-derived cells to produce MSCs. Various culture conditions, including the seeding concentrations of cells and the media used, were investigated. We were able to obtain adherent cell populations after 3 to 5 weeks in our culture conditions from UCB-derived mononuclear cell fractions that had undergone cryopreservation for 0.1 to 5 years. These cells exhibited a fibroblast-like morphology and typical mesenchymal-like immunophenotypes. The results indicate that cryopreserved human UCB fractions can be used as an alternative source of MSCs for experimental and clinical applications as well as for tissue engineering.

Controlled differentiation of stem cells

“…A similar strategy has been applied to isolate clinically applicable mesenchymal precursor cells from hESCs without feeder support and EB formation [79]. MSCs have been derived from a number of different adult tissues, such as bone marrow, deciduous teeth, adipose tissue, umbilical cord blood, and synovium [4,[80][81][82][83]. MSCs have the capability to differentiate into mesodermal lineages, such as adipogenic, osteogenic, chondrogenic, and myogenic cells and can potentially be utilized as a cell source for musculoskeletal tissue regeneration [84,85].…”

Clonal analysis of nestin– vimentin+ multipotent fibroblasts isolated from human dermis

“…Multipotent adult stem cells seem to be almost comparable to embryonic stem cells in respect of their ability to differentiate into various cell types in vitro and in vivo. Adult mesenchymal stem cells have been identified in bone marrow Mackay et al, 1998;Muraglia et al, 2000;Pittenger et al, 1999), skeletal muscle (Asakura et al, 2001;Lee et al, 2000) and adipose tissue (De Ugarte et al, 2003;Zuk et al, 2002), as well as in cord blood (Erices et al, 2003;Lee et al, 2005). Cells isolated from those tissues are usually a heterogeneous mixture of different subpopulations.…”

Stem cells for spine surgery

“…Adult stem cells have more commonly been used in both in vitro and in vivo experimentation due to the ethical concerns regarding embryonic stem cells. MSCs are favored by many scientists due to the ease of cell harvest, isolation, expansion, and preservation [35,[95][96][97] . To date, no reports of immunologic reactions to allogeneic vs autologous cell transplants have been observed, making MSCs a very practical solution for cellular therapy [98,99] .…”