Stojkovic M, Kölle S, Peinl S, Stojkovic P, Zakhartchenko V, Thompson J G, Wenigerkind H, Reichenbach H-D, Sinowatz F, Wolf E
Department of Molecular Animal Breeding and Biotechnology, Hackerstr. 27, D-85764 Oberschleissheim, Germany.
Reproduction. 2002 Jul;124(1):141-53.
Hyaluronic acid (HA) is the main glycosaminoglycan present in follicular, oviductal and uterine fluids. The main functions of HA include dynamic processes that are mediated through interaction with extracellular matrix components, regulation of gene expression, cell proliferation and cell differentiation. HA increases the viscosity of solutions and also has several physiological functions, including regulation of water distribution and water-binding capacity. The addition of 6 mg HA ml(-1) to synthetic oviduct fluid (SOF; SOF-HA) culture medium on day 5 (IVF = day 0) significantly (P < 0.001) increased the viscosity of the medium in comparison with SOF culture medium containing BSA (SOF-BSA). On day 8, rate of blastocyst development in SOF-HA culture medium was significantly (P < 0.05) higher than in SOF-BSA culture medium (38.2 versus 29.3%). The number of trophectoderm cells and the total number of cells of expanded blastocysts cultured in the presence of HA were significantly (P < 0.01) higher in comparison with expanded blastocysts cultured in the presence of BSA (88.9 +/- 7.3 versus 67.6 +/- 3.0 and 130.1 +/- 10.9 versus 104.8 +/- 2.5, respectively). After freezing and thawing, the percentage of day 8 embryos that re-expanded and hatched when cultured with SOF-HA was greater than that of embryos cultured with SOF-BSA (11.3 and 10.5% versus 75.5 and 36.8%, respectively). After thawing, the ATP contents of in vivo-derived, SOF-HA and SOF-BSA expanded blastocysts were similar. The embryos cultured with HA showed less ultrastructural deviation and de-differentiation after freezing and thawing than the embryos cultured with BSA. This study demonstrates that HA improves the developmental capacity of bovine embryos under in vitro conditions and is warranted as a supplement for in vitro production of bovine embryos, particularly if they are to be cryopreserved.
透明质酸（HA）是卵泡液、输卵管液和子宫液中存在的主要糖胺聚糖。HA的主要功能包括通过与细胞外基质成分相互作用介导的动态过程、基因表达调控、细胞增殖和细胞分化。HA可增加溶液的粘度，还具有多种生理功能，包括调节水的分布和水结合能力。在第5天（体外受精=第0天）向合成输卵管液（SOF；SOF-HA）培养基中添加6mg HA/ml（-1），与含牛血清白蛋白的SOF培养基（SOF-BSA）相比，显著（P<0.001）增加了培养基的粘度。在第8天，SOF-HA培养基中囊胚发育率显著（P<0.05）高于SOF-BSA培养基（38.2%对29.3%）。与在牛血清白蛋白存在下培养的扩张囊胚相比，在HA存在下培养的扩张囊胚的滋养外胚层细胞数量和细胞总数显著（P<0.01）更高（分别为88.9±7.3对67.6±3.0和130.1±10.9对104.8±2.5）。冷冻和解冻后，用SOF-HA培养时第8天重新扩张和孵化的胚胎百分比高于用SOF-BSA培养的胚胎（分别为11.3%和10.5%对75.5%和36.8%）。解冻后，体内来源的、SOF-HA和SOF-BSA扩张囊胚的ATP含量相似。与用牛血清白蛋白培养的胚胎相比，用HA培养的胚胎在冷冻和解冻后超微结构偏差和去分化较少。本研究表明，HA可提高体外条件下牛胚胎的发育能力，作为牛胚胎体外生产的补充剂是有必要的，特别是如果它们要被冷冻保存的话。
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