Liu Qin, Wang Liping, Chen Fang, Zhang Yi
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Feb;33(2):202-5.
To study the feasibility of isolation and culture of adipose-derived stem cells( ADSCs) from SD rat adipose tissues subjected to long-term cryopreservation.
We took inguinal fat pads from healthy SD rats. Adipose tissues were stored with 100 m L / L dimethyl sulfoxide( DMSO) combined with 900 m L / L fetal bovine serum( FBS) in liquid nitrogen. Three months later,the adipose tissues were resuscitated for the isolation and culture of ADSCs. The growth status and morphology were observed. The growth curve and cell surface markers CD29,CD45,CD90 of the 3rd passage cells were analyzed respectively by CCK-8 assay and immunocytochemistry. The 3rd passage cells were induced towards adipogenic lineages and osteogenic lineages by different inducers,and the resulting cells were examined separately by oil red O staining and alizarin red staining.
The ADSCs obtained from SD rat adipose tissues subjected to long-term cryopreservation showed a spindle-shape appearance and had a good proliferation ability. The cell growth curve was typical "S " curve.Immunocytochemistry showed that the 3rd passage cells were positive for CD29 and CD90,while negative for CD45. The cells were positive for oil red O staining after adipogenic induction,and also positive for alizarin red staining after osteogenic induction.
The ADSCs can be isolated from SD rat adipose tissues subjected to long-term cryopreservation.
研究从长期冷冻保存的SD大鼠脂肪组织中分离培养脂肪干细胞（ADSCs）的可行性。
取健康SD大鼠腹股沟脂肪垫。脂肪组织用含100 mL/L二甲基亚砜（DMSO）和900 mL/L胎牛血清（FBS）的混合液在液氮中保存。3个月后，复苏脂肪组织用于ADSCs的分离培养。观察细胞生长状态和形态。分别采用CCK-8法和免疫细胞化学法分析第3代细胞的生长曲线及细胞表面标志物CD29、CD45、CD90。用不同诱导剂将第3代细胞诱导分化为脂肪细胞和成骨细胞系，分别采用油红O染色和茜素红染色检测诱导后的细胞。
从长期冷冻保存的SD大鼠脂肪组织中获得的ADSCs呈纺锤形外观，增殖能力良好。细胞生长曲线呈典型的“S”形。免疫细胞化学显示第3代细胞CD29和CD90呈阳性，CD45呈阴性。成脂诱导后细胞油红O染色呈阳性，成骨诱导后茜素红染色也呈阳性。
可从长期冷冻保存的SD大鼠脂肪组织中分离出ADSCs。
