The effect of gestational diabetes on proliferation capacity and viability of human umbilical cord-derived stromal cells

Abstract

Stomal cells derived from Wharton's jelly of human umbilical cord (WJMSCs) are considered as the potential therapeutic agents for regeneration and are getting famous for stem cell banking. Our study aims to evaluate the effects of gestational diabetes on proliferation capacity and viability of WJMSCs. Mesenchymal stromal cells were isolated from Wharton's jelly of human umbilical cords from normal and gestational diabetic (DWJMSCs) mothers. Growth patterns of both types of cells were analyzed through MTT assay and population doubling time. Cell survival, cell death and glucose utilization were estimated through trypan blue exclusion assay, LDH assay and glucose detection assay respectively. Angiogenic ability was evaluated by immunocytochemistry and ELISA for VEGF A. Anti-cancerous potential was analyzed on HeLa cells. DWJMSCs exhibited low proliferative rate, increased population doubling time, reduced cell viability and increased cell death. Interestingly, DWJMSCs were found to have a reduced glucose utilization and anti-cancerous ability while enhanced angiogenic ability. Gestational diabetes induces adverse effects on growth, angiogenic and anti-cancerous potential of WJMSCs.

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“…Gestational diabetes mellitus (GDM) impairs the proliferative capacity of WJMSCs. The key factor involved in GDM is excessive amount of glucose in mother which crosses freely to fetus 7 . In order to achieve maximum therapeutic benefits it is a prerequisite to optimize the culture conditions for cellular growth.…”

“…Many pathological conditions including diabetes mellitus are associated with hyperglycemia 5 and oxidative stress which affects the growth of stem cell 6 . It is reported that WJMSCs derived from diabetic mothers (DWJMSCs) show reduced growth rate in culture conditions as compared to normal mothers' derived WJMSCs (NWJMSCs) 7 . This study is based on finding the optimal glucose concentration for maximum growth of NWJMSCs and DWJMSCs in vitro.…”

“…WJMSCs were isolated by explant culturing method from human umbilical cord of mothers with or without gestational diabetes as previously described by us 7 . Briefly, 5 g cord tissue was cut in to smaller pieces which were subsequently incubated in Dulbecco's modified eagle medium low glucose (DMEM LG)(Sigma Aldrich, USA) with 10 % fetal bovine serum(FBS) (Gibco, Grand Island, NJ) and 100 U/ml penicillin and 100 µg/ml streptomycin (Gibco, Grand Island, NJ).…”

“…Amount of reduced glutathione (GSH) in culture medium after treatment was estimated using method of 7,8 . For this, 0.5 mL culture medium was added in a tube along with 2.0 mL disodium hydrogen phosphate buffer (0.3 M) and 0.25 mL 5,5'-dithiobis-(2-nitrobenzoic acid) or DTNB (0.001 M) (In vitro gen Inc., USA).…”

Crosstalk Between Decreased Anti-Oxidant and Increased Serum Angiogenesis in Renal Cell Carcinomas

“…ELISA was done as reported previously [15,16]. Solid phase sandwich ELISA was done for Using a micro-titer plate reader, absorbance was taken at a wavelength of 450 nm along with 650 nm as a reference value.…”