Expansion of mesenchymal stem cells isolated from pediatric and adult donor bone marrow

Abstract

The enormous plasticity of mesenchymal stem cells (MSCs) suggests an improvement of a standard protocol of isolation and ex vivo expansion for experimental and clinical use. We isolated and expanded MSCs from bone marrow (BM) of pediatric and young adult donors, to analyze the growth kinetic, immunophenotype, telomere length, karyotype during ex vivo expansion. Seventeen BM samples were collected from young adult donors and 8 from pediatric donors. MSCs isolated from two groups showed no morphological differences while their cell growth was strictly related to the donor's age. The MSCs isolated from pediatric donors reached a cumulative PD almost twice as high as MSCs isolated from young adult donors after 112 days (10.2 +/- 1.9 versus 5.5 +/- 3.7). Furthermore, we analyzed the modulation of antigen expression in the MSCs isolated from two groups until 10th passage (77 days) and there was no significant difference between the modulation of antigen expression. In particular, at the first passage, MSCs showed a low contamination of hemopoietic cells which became insignificant in the following passages. There was a high expression of CD90, CD29, CD44 and CD105 and variable and moderate expression of CD166 and CD106 at the start of MSC culture and at each passage during expansion. No chromosomal alteration or evidence of cellular senescence were observed in all analyzed samples. All these data suggest that MSCs can be isolated and expanded from most healthy donors, providing for an autologous source of stem cells.

The cell culture expansion of bone marrow stromal cells from humans with spinal cord injury: implications for future cell transplantation therapy

“…MSC population doublings in non-SCI humans have previously been found to relate to age, where cultures established from paediatric donors proliferated to generate nearly twice as many cells as those derived from young adults within the same time span. 18 In addition, differences in the number of mononuclear cells present in bone marrow biopsies taken from SCI donors with cervical versus thoracic injuries have been reported, where donors with cervical lesions had larger cell counts per ml of marrow. 17 We found that the number of MSCs generated at passage I as a proportion of mononucleated cells originally isolated (that is, the MSC index) was inversely related to age in SCI donors but not non-SCI donors.…”

Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

“…Widely disparate results have been reported with regard to MSC numbers, differentiation potency and aging. A negative correlation between donor age, number and proliferative capacity of MSCs isolated from young and old donors has been demonstrated by several authors (Martin et al 1970, Schneider & Mitsui 1976, Majors et al 1997, D'Ippolito et al 1999, Stenderup et al 2003, Mareschi et al 2006, but widely different results have been obtained with regard to MSC antigen expression. Recently, Stolzing et al (2008) confirmed those already demonstrated in previous studies (D 'Ippolito et al 1999, Mendes et al 2002, Chen et al 2005, Zhang et al 2005, Zheng et al 2007, Kretlow et al 2008, observing a significant age-related change in membrane markers expression levels, an alkaline phosphatase activity, chondrogenic and myogenic differentiation potency decline with donor age in contrast with adipogenic differentiation.…”

Assessment of therapeutic efficacy and fate of engineered human mesenchymal stem cells for cancer therapy

“…H uman mesenchymal stem cells (MSCs) are multipotent cells that can self-renew, proliferate, and differentiate into a variety of cell types (1,2). A significant improvement in understanding MSC biology in recent years has paved the way to their potential clinical use.…”