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Original Article

Development of an effective dissociation protocol for isolating mesenchymal stem cells from bovine intermuscular adipose tissues

초록

영어

Intermuscular fat is essential for enhancing the flavor and texture of cultured meat. Mesenchymal stem cells derived from intermuscular adipose tissues are a source of intermuscular fat. Therefore, as a step towards developing a platform to derive intermuscular fat from mesenchymal stem cells (MSCs) for insertion between myofibrils in cultured beef, an advanced protocol of intermuscular adipose tissue dissociation effective to the isolation of MSCs from intermuscular adipose tissues was developed in cattle. To accomplish this, physical steps were added to the enzymatic dissociation of intermuscular adipose tissues, and the MSCs were established from primary cells dissociated with physical step-free and step-added enzymatic dissociation protocols. The application of a physical step (intensive shaking up) at 5 minutes intervals during enzymatic dissociation resulted in the greatest number of primary cells derived from intermuscular adipose tissues, showed effective formation of colony forming units-fibroblasts (CFU-Fs) from the retrieved primary cells, and generated MSCs with no increase in doubling time. Thus, this protocol will contribute to the stable supply of good quality adipose-derived mesenchymal stem cells (ADMSCs) as a fat source for the production of marbled cultured beef.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Preparation of intermuscular adipose tissues
Harvest of primary cells from intermuscular adipose tissues
Establishment and culture of ADMSCs from intermuscular adipose tissue-derived primary cells
CFU-F assay
Measurement of doubling time
Real-time polymerase chain reaction (PCR)
Statistical analysis
RESULTS
Influence of the addition of physical steps to the enzymatic dissociation protocol on the retrieval of primary cells from intermuscular adipose tissues in cattle
Comparison of marker expression and doubling time of MSCs established from primary cells retrieved from intermuscular adipose tissues using enzymatic dissociation without or with additional physical steps
DISCUSSION
CONCLUSION
REFERENCES

저자정보

  • Jeong Min Lee Department of Applied Animal Science, Kangwon National University, Kustogen, Chuncheon 24341, Korea
  • Hyun Lee Department of Applied Animal Science, Kangwon National University, Chuncheon 24341, Korea
  • Seung Tae Lee Department of Applied Animal Science, Kangwon National University, Kustogen, Chuncheon 24341, Korea

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