Differential expression profiles of lncRNAs and mRNAs during male germ-like cell differentiation in human umbilical mesenchymal stem cells by RNA-seq

Background: Long non-coding RNAs (lncRNAs) are key regulators of various biological processes and crucial for cell development and differentiation. However, their roles in the differentiation of human umbilical mesenchymal stem cells (HUMSCs) into male germ-like cells remain largely unknown. Method: Here, the expression of lncRNAs and mRNAs in undifferentiated HUMSCs and HUMSCs undergoing differentiation into male germ-like cells was analyzed. RNA-sequencing was performed to profile the expression of non-coding RNAs. We analyzed the total expression of lncRNAs/mRNAs at three time points during HUMSC differentiation [day (D)7, D14, and D21]. Result: Expression profiling revealed 110 lncRNAs, 584 mRNAs, and 21 miRNAs common to the three experimental groups during HUMSC male germ-like cell differentiation. The maximum and minimum total overall lncRNA expression occurred on D14 (638) and D21 (283), respectively. The maximum and minimum numbers of up-regulated mRNAs were observed on D21 (2,398) and D7 (2,106), respectively. The maximum and minimum numbers of down-regulated mRNAs were observed on D14 (3,357) and D21 (202), respectively. The expression level of mRNA ENST00000486554 was up-regulated on D7, D14, and D21 after induction. Pathway analysis identified meiotic signaling pathways and nitrogen metabolism as being associated with the differentiation potential of HUMSC male germ-like cells. Non-coding RNA expression profiles significantly differed in HUMSC male germ-like cell differentiation. One mRNA, ENST00000486554, was crucial for differentiation. Conclusions: Our results provide a systematic perspective on the potential functions of non-coding RNAs and novel insights into the complicated regulatory mechanisms underlying the differentiation of HUMSCs into male germ-like cells.

Comparison of reversal of rat pulmonary fibrosis of nintedanib, pirfenidone, and human umbilical mesenchymal stem cells from Wharton’s jelly

Background The present study compared the effects of antifibrotic medications, pirfenidone, and nintedanib, with transplantation of human umbilical mesenchymal stem cells (HUMSCs) in restoring rat pulmonary fibrosis (PF). Methods A stable animal model was established via an intratracheal injection of 5 mg bleomycin (BLM). One single transplantation of 2.5× 107 HUMSCs or initiation of daily oral nintedanib/pirfenidone administration was performed on day 21 following BLM damage. Results Pulmonary function examination revealed that BLM rats exhibited a significant decrease in blood oxygen saturation and an increase in respiratory rates. While no significant improvements were found in BLM rats receiving nintedanib or pirfenidone, those who transplanted with HUMSCs showed a statistical amelioration in blood oxygen saturation and significant alleviation in respiratory rates. Quantification results revealed that a significant reduction in alveolar space and marked increases in substantial cell infiltration and collagen deposition in the left lungs of BLM rats. No significant alteration was observed in BLM rats administered nintedanib or pirfenidone. However, BLM rats transplanted with HUMSCs had a significant recovery in alveolar space and noticeable decreases in cell infiltration and collagen deposition. The inflammatory cell numbers in the bronchoalveolar lavage was increased in the BLM group. While the rats treated with nintedanib or pirfenidone had a lower cell number than the BLM group, a higher cell number was found as compared with the Normal group. In rats transplanted with HUMSCs, the cell number did not differ from the Normal group. Conclusions Transplantation of HUMSCs could effectively treat PF as opposed to the administration of anti-fibrotic drugs with nintedanib or pirfenidone with a significant better result in lung volume, pathological changes, lung function, and blood oxygen saturation.

Comparison of Reversal of Rat Pulmonary Fibrosis of Nintedanib, Pirfenidone and Human Umbilical Mesenchymal Stem Cells from Wharton’s Jelly

Background:The present study compared the effects of antifibrotic medications, Pirfenidone and Nintedanib, with transplantation of human umbilical mesenchymal stem cells (HUMSCs) in restoring rat pulmonary fibrosis (PF).Methods:A stable animal model was established via an intratracheal injection of 5 mg bleomycin (BLM). One single transplantation of 2.5 107 HUMSCs or initiation of daily oral Nintedanib/Pirfenidone administration was performed on Day 21 following BLM damage.Results:Pulmonary function examination revealed that BLM rats exhibited a significant decrease in blood oxygen saturation and an increase in respiratory rates. While no significant improvements were found in BLM rats receiving Nintedanib or Pirfenidone, those who transplanted with HUMSCs showed statistical amelioration in blood oxygen saturation and significant alleviation in respiratory rates. Quantification results revealed that a significant reduction in alveolar space and marked increases in substantial cell infiltration and collagen deposition in the left lungs of BLM rats. No significant alteration was observed in BLM rats administered Nintedanib or Pirfenidone. However, BLM rats transplanted with HUMSCs had a significant recovery in alveolar space and noticeable decreases in cell infiltration and collagen deposition. The inflammatory cell numbers in the bronchoalveolar lavage was increased in the BLM group. While the rats treated with Nintedanib or Pirfenidone had a lower cell number than the BLM group, a higher cell number was found as compared with the Normal group. In rats transplanted with HUMSCs, the cell number did not differ from the Normal group.Conclusions:Transplantation of HUMSCs could effectively treat PF as opposed to the administration of anti-fibrotic drugs with Nintedanib or Pirfenidone with significant better result in lung volume, pathological changes, lung function and blood oxygen saturation.

Comparison of Reversal of Rat Pulmonary Fibrosis of Nintedanib, Pirfenidone and Human Umbilical Mesenchymal Stem Cells from Wharton’s Jelly

Background:The present study compared the effects of antifibrotic medications, pirfenidone and nintedanib, with transplantation of human umbilical mesenchymal stem cells (HUMSCs) in restoring rat pulmonary fibrosis (PF).Methods:A stable animal model was established via an intratracheal injection of 5 mg bleomycin (BLM). One single transplantation of 2.5 × 107 HUMSCs or initiation of daily oral nintedanib/pirfenidone administration was performed on Day 21 following BLM damage.Results:Pulmonary function examination revealed that BLM rats exhibited a significant decrease in blood oxygen saturation and an increase in respiratory rates. While no significant improvements were found in BLM rats receiving nintedanib or pirfenidone, those who transplanted with HUMSCs showed statistical amelioration in blood oxygen saturation and significant alleviation in respiratory rates. Quantification results revealed that a significant reduction in alveolar space and marked increases in substantial cell infiltration and collagen deposition in the left lungs of BLM rats. No significant alteration was observed in BLM rats administered nintedanib or pirfenidone. However, BLM rats transplanted with HUMSCs had a significant recovery in alveolar space and noticeable decreases in cell infiltration and collagen deposition. The inflammatory cell numbers in the bronchoalveolar lavage was increased in the BLM group. While the rats treated with nintedanib or pirfenidone had a lower cell number than the BLM group, a higher cell number was found as compared with the Normal group. In rats transplanted with HUMSCs, the cell number did not differ from the Normal group.Conclusions:Transplantation of HUMSCs could effectively treat PF as opposed to the administration of anti-fibrotic drugs with nintedanib or pirfenidone with significant better result in lung volume, pathological changes, lung function and blood oxygen saturation.

P0390THERAPEUTIC EFFECTS AND SERUM METABOLOMIC VARIATIONS IN FOCAL SEGMENTAL GLOMERULOSCLEROSIS MICE ON TREATMENT WITH HUMAN UMBILICAL MESENCHYMAL STEM CELLS

Background and Aims Focal segmental glomerulosclerosis (FSGS) mediates the kidney podocyte dysfunction and leads to chronic renal diseases worldwide. Current treatment options are limited, owing to the poor understanding of the metabolic pathologic variations and regeneration of FSGS. Mesenchymal stem cells (MSC) offer the better insights to prevent renal injury and could promote the recovery of renal structure and function through complex mechanisms. Due to the safe, feasible, and therapeutic features, we first designed to investigate the role of human umbilical mesenchymal stem cells (HUMSCs) on FSGS mice and explored the potential serum metabolomic variations. Method Nephropathy was induced by adriamycin (ADR) in male Balb/C mice to study FSGS via intravenous administration. 2*106 HUMSCs are provided to treat FSGS model for 3 times. Renal function was measured by urine protein, serum urea nitrogen (BUN) and serum creatinine (SCr) in the study. Serum metabolic profiles from three groups of Balb/C mice were analysed using ultra- performance liquid chromatography coupled with a triple quadrupole-linear ion trap mass spectrometer and a novel mass spectrometry (UPLC-MS/MS) data collection technique. Results The concentrations of SCr and BUN were apparently decreased after HUMSCs injection for three times compared with the FSGS model and body weight from HUMSCs group was improved significantly. A total of 532 differential metabolites were identified between FSGS group and the normal control group. Bile acid dysregulation and valine/leucine/ isoleucine metabolism pathway, especially, N-acetylvaline and L-valine, were positively associated to ADR-induced FSGS. A panel of identified specific metabolites (pyrrole-2- carboxylic acid, urocanic acid and 6-hydroxynicotinic acid) as well as valine/leucine/isoleucine metabolism pathway rather than bile acid metabolism were related to the renal repair after HUMSCs treatment. Conclusion The administration of HUMSCs in FSGS induced ADR provides the promising protection owing to the improvement of metabolomics. Figure 2. The distribution of DiR-HUMSCs in vivo. Figure 3. Kidney pathology using periodic acid-Schiff (PAS) and Masson staining. Figure 4. Metabolome profiling analysis and Venn diagram for comparisons of numbers from each group. Figure 5. Metabolic pathway analysis results from FSGS + Saline and FSGS + HUMSCs groups. Figure S1. The quality control (QC) of serum metabolomics was designed and performed as follows. Figure S2. Metabolic pathway analysis results from FSGS + Saline and Normal + Saline groups. Figure S3. Metabolic pathway analysis results from FSGS + HUMSCs and Normal + Saline groups.