Functionally heterogeneous human satellite cells identified by single cell RNA sequencing.

Although heterogeneity is recognized within the murine satellite cell pool, a comprehensive understanding of distinct subpopulations and their functional relevance in human satellite cells is lacking. We used a combination of single cell RNA sequencing and flow cytometry to identify, distinguish, and physically separate novel subpopulations of human PAX7+ satellite cells (Hu-MuSCs) from normal muscles. We found that, although relatively homogeneous compared to activated satellite cells and committed progenitors, the Hu-MuSC pool contains clusters of transcriptionally distinct cells with consistency across human individuals. New surface marker combinations were enriched in transcriptional subclusters, including a subpopulation of Hu-MuSCs marked by CXCR4/CD29/CD56/CAV1 (CAV1+). In vitro, CAV1+ Hu-MuSCs are morphologically distinct, and characterized by resistance to activation compared to CAV1- Hu-MuSCs. In vivo, CAV1+ Hu-MuSCs demonstrated increased engraftment after transplantation. Our findings provide a comprehensive transcriptional view of normal Hu-MuSCs and describe new heterogeneity, enabling separation of functionally distinct human satellite cell subpopulations.

Regressing an optimal variable to scale glomerular filtration rate: more variations in glomerular filtration rate are explained.

BACKGROUND: To compare glomerular filtration rate (GFR) among individuals, GFR is usually scaled to body surface area (BSA) based on the ratio method, which has been debated for its accuracy in recent years. Reference to the BSA as a normalization standard is the most common method currently in use but has limitations. This study was designed to a better variable to scale GFR. METHODS: We measured 99mTc- diethylene triamine pentaacetic acid plasma clearance (uncorrected GFR, uGFR) for 322 healthy adults who were enrolled according to the SENIEUR protocol. The individuals were randomly grouped into A and B for regressing and validating the optimal variable, respectively. Nonlinear regression was performed against uGFR, and the selected independent variables were body weight, height, age, and sex. RESULTS: Among several tested models, the regression coefficients of weight-age formula (W-A) were in narrower 95% confidence interval (CI). The coefficient of determination of the regression line between W-A and uGFR, as an indicator to explain the percentage of variations of GFR, was higher than that of other variables in both groups. The coefficient of determination of the regression line between W-A and uGFR was 0.571, which was higher than that of BSA (0.203) or TBW (0.241). CONCLUSION: The index variable, based on both body weight and age, has a better statistical relationship to uGFR and is a better variable to scale GFR in adults.

Mesenchymal stem cells protect podocytes from apoptosis induced by high glucose via secretion of epithelial growth factor.

INTRODUCTION: The apoptosis and subsequent injury of podocytes plays a pathogenic role in diabetic nephropathy (DN). Mesenchymal stem cells (MSCs) are promising therapeutic cells for preventing apoptosis and reducing cellular injury. Our previous study found that MSCs could protect kidneys from diabetes-induced injury without obvious engraftment. So we evaluated the effects of human adipose-derived MSCs (hAd-MSCs) on podocytic apoptosis and injury induced by high glucose (HG) and the underlying mechanisms. METHODS: We used flow cytometry, Western blot and confocal fluorescence microscopy to study podocytic apoptosis and injury induced by HG at 24 hours, 48 hours, and 72 hours in the presence or absence of MSC-conditioned medium (CM). An antibody-based cytokine array was used to identify the mediating factor, which was verified by adding the neutralizing antibody (NtAb) to block its function or adding the recombinant cytokine to the medium to induce its function. RESULTS: hAd-MSC-CM reduced podocytic apoptosis in a dose-dependent manner, decreased the expression of podocytic cleaved caspase-3, and prevented the reduced expression and maintained the normal arrangement of podocytic synaptopodin and nephrin. However, human embryonic lung cell (Wi38)-CM failed to ameliorate podocytic apoptosis or injury. Twelve cytokines with concentration ratios (MSC-CM/Wi38-CM) >10-fold were identified. Epithelial growth factor (EGF) was singled out for its known ability to prevent apoptosis. Recombinant human EGF (rhEGF) prevented podocytic apoptosis and injury similarly to hAd-MSC-CM but, upon blockade of EGF, the beneficial effect of hAd-MSC-CM decreased dramatically. CONCLUSIONS: hAd-MSCs prevent podocytic apoptosis and injury induced by HG, mainly through secreting soluble EG.