Figure 2.

hMPCs exhibited limited capacity for serine/glycine biosynthesis.

A) Heatmap of genes involved in serine, glycine, and one-carbon metabolism based on RNA-seq data from hMPCs after 5 days of culture in serine/glycine replete (1000 μM) or restricted conditions.

B) Immunoblot for PHGDH, PSAT1, PSPH, SHMT1, and SHMT2 protein normalized to α-TUBULIN for quantification in hMPCs that had been serine/glycine restricted for 5 days.

C) Glucose uptake by hMPCs after 5 days of culture in serine/glycine replete or restricted conditions.

D) Live cell count of hMPCs cultured in serine/glycine replete media or serine/glycine restricted media and varying doses of glucose. ∗p < 0.05 and ∗∗p < 0.01 relative to serine/glycine containing control. Data are expressed as mean ± SD.

E) Percent mass isotopomer distribution of [U–¹³C₆]-glucose-derived serine and glycine in hMPCs cultured in serine/glycine replete (1000 μM) or restricted media for 5 days followed by 48 h in similar media but containing [U–¹³C₆]-glucose. Data are expressed as mean ± SD. ∗∗p < 0.01 and ∗∗∗p < 0.001.

All the experiments were repeated with hMPCs derived from the same 5 donors. P values or marks of significance are indicated in the appropriate graphs.