| 摘要: |
| 【目的】 比较体外条件下不同分离山羊卵巢间质细胞(ovarian interstitial cells)方法的效果。【方法】 采用热消化法、冷消化法、单一酶消化法（热消化法，用Ⅰ型、Ⅳ型、Ⅺ型胶原酶和2.5 g/L 胰蛋白酶+0.2 g/L EDTA）及2步酶消化法分离山羊卵巢间质细胞。【结果】 热消化法得到的细胞数量较冷消化法多,且试验所用时间缩短，但细胞活率降低。单一酶热消化法中，以1 g/L Ⅺ型胶原酶效果最佳，适合体外培养。2步酶消化法以先胰蛋白酶后Ⅺ型胶原酶组的效果最佳，卵巢组织消化时间及体外培养首次换液时间均较短，适合于体外培养。【结论】 采用1 g/L Ⅺ型胶原酶单一酶热消化法或先胰蛋白酶后Ⅺ型胶原酶2步酶消化法，分离山羊卵巢间质细胞的效果均较好，可作为试验用卵巢间质细胞的理想分离方法。 |
| 关键词: 山羊 卵巢间质细胞 细胞分离 细胞培养 |
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| 基金项目:陕西省重点攻关项目“干细胞与家畜胚胎工程示范”（2006KZ05-G1） |
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| Comparison of the methods for isolating goat ovarian interstitial cells |
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QUAN Fu sheng YU Yuan SU Hui min HUA Song ZHENG Yue mao LIU Jun ZHANG Yong
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| Abstract: |
| 【Objective】 Different methods for isolating goat ovarian interstitial cells (OICs) were compared in order to obtain adequate cells for in vitro culture in this study.【Method】 The effect of isolation was investigated in different temperature(dissociation at 37 ℃ and at 4 ℃ respectively);methods of isolating goat OICs were compared by a single enzyme digestion treatment (collagenase typeⅠ,Ⅳ, Ⅺ and trypsinase + EDTA,at 37 ℃) and two-step enzymatic digestion treatment.【Result】 The results showed that dissociation at 37 ℃ could obtain adequate cells and save an amount of time than at 4 ℃,but remained low cell viability rates;as a single enzyme digestion treatment,1 g/L collagenase type Ⅺ was most suitable for culture in vitro;as a two step enzymatic treatment,Group Ⅱ(trypsinase 15 min first,then collagenase type Ⅺ 15 min) could obtain adequate cells with high viability rates,the digestion time and the time of first exchange of the total volume medium were shortened,and also suitable for culture in vitro.【Conclusion】 The two methods of isolating goat OICs were satisfactory for experimental use by 1 g/L collagenase type Ⅺ digestion treatment and two-step enzymatic treatment (trypsinase 15 min first,then collagenase type Ⅺ 15 min) at 37 ℃. |
| Key words: goat ovarian interstitial cell cell isolation cell culture |
