A procedure has been developed for isolating protoplasts from prothalli of Ceratopteris richardii which can be cultured and are capable of regeneration. Protoplasts were isolated from 2-week-old gametophytes in a medium containing wall-digesting enzymes in 0.5 M sucrose, followed by purification of the released protoplasts by floating them up into a 0.5 M sorbitol layer. Regeneration occurred over a period of 10-24 days, and, under optimal osmotic conditions, followed the developmental pattern seen during spore germination, in that the first division gave rise to a primary rhizoid. Thus, prothallial protoplasts are comparable to germinating spores as suitable models for studies of developmental polarity in single cells. As in germinating spores, the polarity of development in regenerating protoplasts is influenced by the vectors of gravity and unilateral light. However, the relative influence of light in fixing this polarity is greater in regenerating protoplasts, while in germinating spores, the influence of gravity is greater.

中文翻译：

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重力和光控制的再生蕨原生蕨的角膜前缘细胞的原生质体的发育极性。

已经开发了一种用于从番茄角鲨的原球藻中分离原生质体的方法，该原生质体可以被培养并且能够再生。在含有0.5M蔗糖的消化酶的培养基中，从2周龄的配子体分离原生质体，然后通过将释放的原生质体上浮到0.5M山梨醇层中来纯化释放的原生质体。再生发生在10-24天的时间内，并且在最佳渗透条件下，遵循孢子萌发过程中所见的发育模式，因为第一次分裂产生了初级根状茎。因此，作为研究单细胞发育极性的合适模型，前藻原生质体与发芽孢子相当。就像在萌芽孢子中一样 原生质体再生的发育极性受重力和单侧光矢量的影响。然而，光在固定该极性方面的相对影响在再生原生质体中更大，而在发芽孢子中，重力的影响更大。