Department of Neuroscience, Medical College of Georgia, Augusta, GA 30912, USA.
J Tissue Eng Regen Med. 2011 Feb;5(2):130-7. doi: 10.1002/term.297.
Successful in vitro differentiation of spermatogenic cells into spermatids appears to offer extremely attractive potential for the treatment of impaired spermatogenesis and male infertility. Experimental evidence indicates that biocompatible polymers may improve in vitro reconstitution and regeneration of tissues of various origins. Here, we fabricated highly porous biodegradable poly(D,L-lactic-co-glycolic acid) or PLGA co-polymer scaffolds by combining the gas-foaming and salt-leaching methods, using ammonium bicarbonate as a porogen, which allowed us to generate polymer scaffolds with a high density of interconnected pores of 400-500 µm in average diameter, concomitant with a high malleability to mould a wide range of temporal tissue scaffolds requiring a specific shape and geometry. The PLGA scaffolds were biocompatible and biodegradable, as evidenced by the fact that they survived almost 3 month long subcutaneous xenografting into immunodeficient host mice and became easily destroyable after recovery. Immature rat testicular cells that were seeded onto the surface of the scaffold exhibited about 65% seeding efficiency and up to 75% viability after 18 days in culture. Furthermore, our scaffolds enhanced the proliferation and differentiation of spermatogenic germ cells to a greater extent than conventional in vitro culture methods, such as monolayer or organ culture. Taken together, an implication of the present findings is that the PLGA-based macroporous scaffold may provide a novel means by which spermatocytes could be induced to differentiate into presumptive spermatids.
体外成功分化生精细胞为精细胞,为治疗受损的生精和男性不育提供了极具吸引力的潜在可能。实验证据表明,生物相容性聚合物可能改善各种来源组织的体外重建和再生。在这里,我们使用碳酸氢铵作为造孔剂,结合气体发泡和盐浸出方法,制造了高度多孔的可生物降解的聚(D,L-乳酸-共-乙醇酸)或 PLGA 共聚物支架,这使得我们能够生成聚合物支架具有平均直径为 400-500 µm 的高密度互连孔,同时具有高度的可模塑性,可以模制需要特定形状和几何形状的广泛的临时组织支架。PLGA 支架具有生物相容性和可生物降解性,事实证明,它们在免疫缺陷宿主小鼠的皮下异种移植中存活了近 3 个月,并且在恢复后很容易被破坏。接种在支架表面的未成熟大鼠睾丸细胞在培养 18 天后,其接种效率约为 65%,存活率高达 75%。此外,我们的支架比传统的体外培养方法(如单层或器官培养)更能促进生殖细胞的增殖和分化。总之,本研究结果表明,基于 PLGA 的大孔支架可能为诱导精母细胞分化为精原细胞提供了一种新方法。