Isolation of mesenchymal stem cells from G-CSF-mobilized human peripheral blood using fibrin microbeads

Abstract

Adult mesenchymal stem cells (MSC) that are able to differentiate into various mesenchymal cell types are typically isolated from bone marrow, but their significant presence in human peripheral blood (PB) is controversial. Fibrin microbeads (FMB) that bind matrix-dependent cells were used to isolate MSC from the mononuclear fraction of mobilized PB of adult healthy human donors treated with a granulocyte colony-stimulating factor. Isolation by plastic adherence resulted in a negligible number of MSC in all samples tested, whereas FMBbased isolation yielded spindle-shaped cell samples that could further expand on plastic or on FMB in eight out of the 11 samples. The yield of these cells at days 17-18 after the harvest was B0.5% of the initial cell number. The isolated cells were grown on plastic and characterized by FACS analysis and immunohistochemistry for specific markers. Following culturing and first passage, the FMBisolated cells stained positive for mesenchymal stromal cell markers CD90 and CD105, expressed vimentin and fibronectin and were negative for hematopoietic markers CD45 and CD34. These cells could differentiate into osteoblasts, adipocytes and chondrocytes. This study indicates that FMB may have special advantage in isolating MSC from sources such as mobilized PB, where the number of such cells is scarce.

Human peripheral blood derived mesenchymal stem cells demonstrate similar characteristics and chondrogenic differentiation potential to bone marrow derived mesenchymal stem cells

“…8 It is worth noting that in few studies, the culture of PB-MSCs has been only occasionally achieved and not always replicable. 23 On the contrary, other studies have demonstrated that MSCs can be isolated from PB repeatedly, however their limitations remains in the low numbers of cells obtained.…”

“…This involves the hyper stimulation of BM production using granulocyte colony-stimulating factor (G-CSF). 8 This, however, results in the production of immature progenitor cells, for example, blasts, rather than the true multipotential MSCs. 11 In addition, it has also been demonstrated that this technique does not selectively produce MSCs but rather a mix of MSCs, hematopoietic stem cells and, other immature progenitor cells.…”

“…7 More recently, MSCs derived from peripheral blood (PB-MSCs) have also been reported, although many researchers have argued that the existence of these cells circulating in adequate numbers remains low. 8,9 According to Roufosse et al, 10 when tissue is damaged, MSCs are transported from BM to the damaged site via the circulating PB to promote regeneration. Using this principle, researchers have demonstrated that MSCs can be increased in the peripheral circulation by a technique known as ''blood mobilization.''…”

Sangue periférico como fonte de células para terapia celular

“…32 Desde então, inúmeros relatos têm surgido na literatura demonstrando a presença, no sangue periférico, de células com capacidade de se diferenciarem em outras células no tecido conjuntivo. [33][34][35][36][37][38] Tondreau e col. 34 testaram o potencial do sangue periférico mobilizado com fator de crescimento como fonte de células-tronco mesenquimais. Os autores demonstraram que CTM podem ser facilmente isoladas a partir da fração de células CD133+ do sangue périférico mobilizado.…”

The contribution of bone marrow-derived cells to the human adipocyte pool

“…37,38 Another type of multipotent stem cells are the MSCs which have the capacity to develop into functional cells of the mesenchymal lineage e.g., osteocytes, chondrocytes and adipocytes. 46,47 As MSCs can be found in both BM and PBSC 48,49 it could be speculated that these cells may constitute "adipogenic" BMDCs. Based on data from murine models, additional cell sources could be endothelial cells.…”