| 摘要: |
| 采用0.1% Ⅺ型胶原酶和2.5 g/L胰蛋白酶0.2 g/L EDTA，分别用热消化法和冷消化法消化分离家兔卵巢间质细胞，以探讨其体外培养条件和生长特点。结果表明，无论用热消化法还是冷消化法，0.1% Ⅺ型胶原酶的消化效果均好于2.5 g/L胰蛋白酶0.2 g/L EDTA的消化效果，前者的细胞活率在90%以上，可用于体外培养；经74 μm滤网过滤后的卵巢组织分离培养出梭形和不规则形的间质细胞，且2种类型的细胞能在同一培养体系中贴壁生长。说明0.1% Ⅺ型胶原酶可用于家兔卵巢间质细胞的分离。 |
| 关键词: 卵巢间质细胞 分离培养 形态观察 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目(39770544) |
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| In vitro culture and morphological observation of ovarian interstitial cells in rabbit |
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| Abstract: |
| The primary goal of this research is to establish the conditions that the ovarian interstitial cells (OICs) can grow well in vitro and observe OICs' growth and morphology characteristics.We use different dissociating solutions (collagenase type Ⅺ and trypsin-EDTA) and methods (dissociation at 37 ℃,20-30 min and at 4 ℃,4-5 h respectively) to derive OICs from the treated ovarian tissue.Then the OICs are cultured in a humidified 95% air-5% CO2 incubator at 37 ℃ supplemented insulin.We find the best dissociated condition is using collagenase at 37 ℃ for 30 min or so.The cell viability is more than 90% with typan blue stain.The OICs are isolated with 74 μm filter.There are two types of cells,one type is shuttle like,the other is irregular.The two types of cells are co-cultured under the same condition and anchored growing on tissue-culture dishes.As a result of our experiments,we conclude that using the 0.1% collagenase type Ⅺ is the best way to isolate the OICs. |
| Key words: OICs isolation and culture morphological observation |
