In vitromeasurement of postnatal changes in proliferating satellite cell frequency during rat muscle growth
In vitromeasurement of postnatal changes in proliferating satellite cell frequency during rat muscle growth
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大鼠肌肉生长过程中增殖卫星细胞频率的出生后变化的体外测量
DOI：
10.1111/j.1740-0929.2009.00734.x
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发表时间：
2011
影响因子：
2
通讯作者：
R
中科院分区：
文献类型：
--
作者：
Suzuki;T.;Takaishi;H.;Sakata;T.;Do;M. Q.;Hara;M.;Sato;A.;Mizunoya;W.;Nishimura;T;Hattori;A.;Ikeuchi;Y.;and Tatsumi;R
卫星细胞是在出生后骨骼肌中发现的常驻肌源性干细胞，在出生后早期发育过程中最为丰富，此后在小鼠和大鼠中的频率急剧下降至成年水平。因此，出生后卫星细胞有丝分裂活动的变化是进一步了解肌肉生长策略的重要方面。然而，在大型肉类生产动物中，传统的活体增殖试验可能不太现实，因为它需要在每个年龄段的宰杀时间内注射大剂量的诱变核苷(IP)、~3H标记的胸腺嘧啶核苷或溴脱氧尿苷(BrdU)。我们在目前的先导性研究中报告，在早期培养的大鼠中，可以通过体外BrdU掺入试验来评估卫星细胞的活体增殖活性。取上后肢和背部肌肉制备卫星细胞，培养24 h，最后2 h加入BrdU，常规免疫细胞化学检测掺入BrdU的细胞百分率。结果表明，在出生后3个月左右，增殖卫星细胞频率迅速下降到成体水平，并与出生后第2天至第11个月的Thein vivoBrdU注射法的测量结果高度相关。通过结合肌肉活检技术，该方法可能进一步适用于大型家畜，使同一生长动物的年龄间隔取样成为可能。
Satellite cells, resident myogenic stem cells found in postnatal skeletal muscle, are most abundant during early postnatal development and sharply decline in frequency thereafter to adult levels in mice and rats. Therefore, postnatal changes in satellite cell mitotic activities are important aspects for further understanding a muscle growth strategy. In large meat‐production animals, however, the traditionalin vivoproliferation assay may be less realistic because it requires intra‐peritoneal (ip) injection of huge dosage of mutagenic nucleosides,3H‐labeled thymidine or bromodeoxyuridine (BrdU), at each age‐time of sacrifice. We report in the present pilot study using rats thatin vivoproliferation activity of satellite cells can be evaluated by anin vitroBrdU‐incorporation assay in early cultures. Briefly, satellite cells were prepared from upper hind‐limb and back muscles and maintained for 24 h with imposing by BrdU addition for the last 2 h, followed by the regular immunocytochemistry for determining BrdU‐incorporated cell percentage. Thisin vitroassay demonstrated a rapid decrease in proliferating satellite cell frequency to the adult level during about 3‐month period after birth, and yielded a high correlation to the measurements by thein vivoBrdU ip‐injection method during the postnatal period examined from day‐2 to month‐11. Thein vitroproliferation assay may be further adaptable for large domestic animals by the combination with a muscle biopsy technique that enables age‐interval sampling from the same growing animals.