In vitro 방법에 의한 근육줄기세포(muscle satellite cells)의 형질 변화 연구
Characterization of muscle satellite cells in vitro
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근육의 줄기세포라고 알려진 근육위성세포(satellite cells, SCs)는 근육의 성장과 발달 및 구조적, 대사적 형질 형성에 중요한 역할을 한다. 본 연구는 근육에서 추출한 SCs의 in vitro분화 성상과 형질을 분석함으로써 SCs가 근육의 성장 및 재생시 형질 형성에 미치는 영향을 구명하기 위해 수행되었다. 6주령 자돈의 Semitendinosus(ST) 근육의 적색근(red semitendinosus; RST)과 백색근(white semitendinosus; WST)에서 SCs를 추출한 후 in vitro 상에서 배양하여 SCs의 증식율, 분화율, 근육 구조단백질인 myosin heavy chain (MyHC) isoforms의 발현 성상을 분석하였다. 추출된 SCs의 숫자는 WST(63 ± 2 × 103 cells/g)보다 RST(79 ± 2 × 103 cells/g) 에서 높게 (P<0.01) 나타났다. Cell doubling time은 RST에서 추출된 SCs가 WST보다 낮았고, 분화도 빨라 WST보다 먼저 근섬유를 형성하였으나, 분화 24h 후에는 WST 유래 SCs의 분화율이 RST보다 높게 나타났다(P<0.05). 하지만, 분화 48h 후 RST와 WST의 SCs간 분화율은 차이가 없는 것으로 나타났다. MyHC type I isoform의 발현은 RST 유래 SCs에서 WST에 비해 높았으나 (P<0.01), type II isoform은 WST 유래 SCs에서 높게 나타났다 (P<0.05). 이 결과는 in vitro에서 배양된 SCs 에서 분화한 근섬유의 성장패턴 및 형질이 유래 근육의 형질을 그대로 반영한다는 것을 보여주는데, 이는 근육 내에 존재하는 SCs는 유래 근육의 형질을 기억하고 있거나, 특정 형질로만 분화되도록 정해져 있다는 것을 의미한다.
Abstract
Muscle satellite cells (SCs), also known as muscle stem cells, have critical roles in growth, development and repair of skeletal muscle. This study was performed to characterize the growth and differentiation performance and phenotypic adaptation of SCs cultured in vitro. To this end, we isolated red and white portion of semitendinosus (ST) muscles and analyzed proliferation and differentiation rate, and myosin heavy chain (MyHC) isoform expression. Cell yield of RST was higher (P < 0.01) than that of WST. SCs from RST showed faster (P < 0.05) doubling time compared with those from WST. At 24 h differentiation, WST showed higher fusion index than RST but there was no difference in fusion index between WST and RST after 48 h differentiation. SCs from RST expressed mostly type I MyHC expression but those from WST expressed mostly type IIB and IIA MyHC. These results indicate that SCs cultured in vitro may remember the phenotype of their muscle of origin or destined to develop into a specific phenotype.
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