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A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells
Published on: August 7, 2017
Biological characterization of sheep kidney-derived mesenchymal stem cells
Meng Ji1, Chunyu Bai1, Lu Li2
1Department of Animal Genetic Resources, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R. China.
Abstract:
The aim of the present study was to isolate, culture and characterize sheep metanephric mesenchymal stem cells (MMSCs). The MMSCs were isolated from the kidney tissue of six-week-old sheep fetus. This study investigated whether primary MMSCs could be grown for 26 passages and expressed Oct-4, which is involved in the self-renewal of undifferentiated pluripotent stem cells. The MMSCs also expressed the renal lineage marker gene PAX2, and mesenchymal cell marker genes CD44, FN1 and VIM. Expression of these genes was detected using immunofluorescence and reverse transcription-polymerase chain reaction assays. Additionally, we observed that the MMSCs are able to differentiate into adipocyte, hepatocyte and chondrocyte cells. Karyotype analyses showed that these cells were 95% diploid and thus differentiated. These results indicate that the MMSCs obtained from sheep fetuses possessed certain characteristics of multipotent stem cells. Therefore, MMSCs may potentially offer utility for tissue engineering and cellular transplantation therapy, and further studies are required to investigate these uses.
Insights
Researchers isolated and cultured sheep metanephric mesenchymal stem cells (MMSCs) from fetal kidney tissue. These MMSCs demonstrated multipotent stem cell characteristics, showing potential for regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Regenerative Medicine
- Sheep Fetal Tissue Research
Background:
- Mesenchymal stem cells (MSCs) are crucial for tissue regeneration.
- Investigating ovine MSCs offers insights into translational applications.
- Characterizing novel stem cell sources is vital for therapeutic development.
Purpose of the Study:
- To isolate, culture, and characterize ovine metanephric mesenchymal stem cells (MMSCs).
- To assess the self-renewal capacity and pluripotency markers of MMSCs.
- To evaluate the differentiation potential and genetic stability of MMSCs.
Main Methods:
- Isolation of MMSCs from six-week-old sheep fetal kidney tissue.
- Culture expansion up to 26 passages.
- Gene expression analysis (Oct-4, PAX2, CD44, FN1, VIM) via immunofluorescence and RT-PCR.
- In vitro differentiation assays (adipogenesis, hepatogenesis, chondrogenesis).
- Karyotype analysis for diploidy assessment.
Main Results:
- Successful isolation and culture of MMSCs from sheep fetal kidneys.
- MMSCs expressed Oct-4 (pluripotency marker) and PAX2 (renal marker), along with mesenchymal markers (CD44, FN1, VIM).
- MMSCs exhibited multipotency, differentiating into adipocytes, hepatocytes, and chondrocytes.
- Karyotype analysis confirmed 95% diploidy, indicating genomic stability.
Conclusions:
- Ovine MMSCs possess characteristics of multipotent stem cells.
- These cells demonstrate potential for tissue engineering and cell transplantation therapies.
- Further research is warranted to explore the therapeutic applications of MMSCs.
