Riley C B, Archer F J, Bailey J V
Department of Veterinary Anesthesiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.
Am J Vet Res. 1996 Aug;57(8):1118-23.
To compare the effects of different commercial nutrient media and sera on protein synthesis and maintenance of cellular density in cultures of the equine superficial digital flexor tendon (SDFT).
8 healthy 2- to 4-year-old horses.
First Dulbecco's modified Eagle's medium, Ham's F12 nutrient mixture, RPMI 1640 medium, minimum essential medium with Earle's salts, minimum essential medium with Hanks' salts, and a Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture with 10% fetal bovine serum (FBS) were compared. Then FBS, fetal equine serum, and donor horse serum, each at 5, 10, and 15% in RPMI 1640 medium, were compared. Explants were cultured in roller bottles at 37 C and aerated (50% O2/45% N2/5% CO2) daily. Rates of [3H]-proline incorporation were used as a measure of the rates of total protein and collagen synthesis on days 13 and 28. Matrix cellular density of explants at days 14 and 28 was measured by computerized image analysis.
Equine SDFT explants were cultured in all media for up to 4 weeks. Proline incorporation was greatest in Ham's F12 nutrient mixture and in RPMI 1640 medium, with the concentration of proline in medium correlating to the in vitro response. Total proline incorporation was greater in 15% FBS than in 5 or 10% FBS. Other differences among sera were not detected. Matrix cell density in 15% donor horse serum was equivalent to that in uncultured controls and higher than that in most other sera at week 2.
The in vitro SDFT culture system described may be used in future studies to enhance knowledge of the biological and biochemical characteristics of intrinsic tendon healing.
比较不同商业营养培养基和血清对马浅屈肌腱（SDFT）培养物中蛋白质合成及细胞密度维持的影响。
8匹健康的2至4岁马。
首先比较了杜氏改良 Eagle 培养基、哈姆氏F12营养混合物、RPMI 1640培养基、含厄尔氏盐的最低必需培养基、含汉克斯氏盐的最低必需培养基以及含10%胎牛血清（FBS）的杜氏改良 Eagle 培养基/哈姆氏F12营养混合物。然后比较了RPMI 1640培养基中分别含5%、10%和15%的FBS、胎马血清和供体马血清。外植体在滚瓶中于37℃培养，每天通气（50% O₂/45% N₂/5% CO₂）。在第13天和第28天，[³H]-脯氨酸掺入率用作总蛋白和胶原蛋白合成速率的指标。通过计算机图像分析测量第14天和第28天外植体的基质细胞密度。
马SDFT外植体在所有培养基中培养长达4周。脯氨酸掺入在哈姆氏F12营养混合物和RPMI 1640培养基中最高，培养基中脯氨酸浓度与体外反应相关。15% FBS中的总脯氨酸掺入量高于5%或10% FBS。未检测到血清之间的其他差异。在第2周时，15%供体马血清中的基质细胞密度与未培养对照相当，且高于大多数其他血清中的密度。
所描述的体外SDFT培养系统可用于未来研究，以增进对肌腱内在愈合的生物学和生化特性的了解。
