Comparative analysis of human UCB and adipose tissue derived mesenchymal stem cells for their differentiation potential into brown and white adipocytes

Abstract

The differentiation potential of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) into brown and white adipocytes in comparison to Adipose tissue derived MSCs (AD-MSCs) were investigated in order to characterize their potency for future cell therapies. MSCs were isolated from ten UCB samples and six liposuction materials. MSCs were differentiated into white and brown adipocytes after characterization by flow cytometry. Differentiated adipocytes were stained with Oil Red O and hematoxylin/eosin. The UCP1 protein levels in brown adipocytes were investigated by immunofluoresence and western blot analysis. Cells that expressed mesenchymal stem cells markers (CD34-, CD45-, CD90+ and CD105+) were successfully isolated from UCB and adipose tissue. Oil Red O staining demonstrated that white and brown adipocytes obtained from AD-MSCs showed 85 and 61% of red pixels, while it was 3 and 1.9%, respectively for white and brown adipocytes obtained from UCB-MSCs. Fluorescence microscopy analysis showed strong uncoupling protein 1 (UCP1) signaling in brown adipocytes, especially which were obtained from AD-MSCs. Quantification of UCP1 protein amount showed 4- and 10.64-fold increase in UCP1 contents of brown adipocytes derived from UCB-MSCs and AD-MSCs, respectively in comparison to undifferentiated MSCs (P < 0.004). UCB-MSCs showed only a little differentiation tendency into adipocytes means it is not an appropriate stem cell type to be differentiated into these cell types. In contrast, high differentiation efficiency of AD-MSCs into brown and white adipocytes make it appropriate stem cell type to use in future regenerative medicine of soft tissue disorders or fighting with obesity and its related disorders.

Melatonin induces fat browning by transdifferentiation of white adipocytes andde novodifferentiation of mesenchymal stem cells

“…59 In this study, we report that in vitro melatonin treatment increases UCP1 expression in MSCdifferentiated adipocytes, suggesting increased brown-type adipogenesis in human and rat-treated MSCs. Another study that compared the white and brown human adipogenesis of MSCs and umbilical cord stem cells showed no differences in the expression of UCP1 between MSCs and differentiated white adipocytes 60 , agreeing with the data observed in MSCs and Differentiated adipocytes from inguinal WAT of ZDF rats and human lipoaspirates. Various studies in humans 61 and rodents 59 revealed that MSCs and preadipocytes involved in adipogenesis showed increased expression of thermogenic genes such as UCP1, indicating their thermogenic potential.…”

“…71,72 We also observed that the expression of CITED1 and PGC1-α was significantly higher in MSCs derived from beige deposits of the inguinal fat pad than in white ones, which suggests a greater commitment of MSCs with the differentiation of thermogenic beige adipocytes in the bMSCs than WMSCs. This could be explained by the epigenomic memory found in beige adipocytes, 46 which could be present in BAT and explain similar data obtained in mice 73 and human studies 60,74,75 in which derived MSCs from BAT were found to express higher levels of thermogenic proteins. A long-term cold exposure study in mice revealed that de novo beige biogenesis was favored from adipogenic precursor cells, 76 suggesting that the beige biogenesis mechanism, either by darkening of pre-existing mature adipocytes or de novo differentiation, also depends on the frequency and duration of stimuli such as cold.…”

“…Human MSCs were isolated from lipoaspirates' adipose tissue. Human abdominal fat was obtained from healthy donors (n = 8) undergoing liposuction plastic surgery (range of age [44][45][46][47][48][49][50][51][52][53][54][55][56][57][58][59][60][61]. All samples used in this study were collected with informed consent and Institutional Review Board approval (ethic permission number: 02/022010 Hospital Virgen de la Victoria, Málaga, Spain).…”

Studying Brown Adipose Tissue in a Human in vitro Context

“…such as bone marrow, blood, and AT (50). MSCs isolated from human AT (termed human multipotent adipose-derived stem (hMADS) cells) could be differentiated to BA-like cells when stimulated with a PPARγ agonist on top of an adipogenic cocktail (51), and with the addition of BMP7 (52). Similarly, bone marrow-derived MSCs may also be able to differentiate to BAlike cells when overexpressing PPARGC1A (53).…”

“…Multipotent mesenchymal stem cells (MSCs) are adult stem cells able to differentiate into a range of mesodermal cell types including adipocytes, and can be isolated from several tissues such as bone marrow, blood, and AT ( 50 ). MSCs isolated from human AT (termed human multipotent adipose-derived stem (hMADS) cells) could be differentiated to BA-like cells when stimulated with a PPARγ agonist on top of an adipogenic cocktail ( 51 ), and with the addition of BMP7 ( 52 ). Similarly, bone marrow–derived MSCs may also be able to differentiate to BA-like cells when overexpressing PPARGC1A ( 53 ).…”