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Ｂｅａｇｌｅ乳狗（５－１０日龄）肾块用热消化法制成细胞批放液氮保存，检定合格后做下列试验：（１）复苏培养长满单层后更换维持液，其细胞能维持４天以上，克服了常规消化培养细胞维持时间短（３６小时）的缺陷。（２）传代１４－２株病毒时，１代病毒的滴度即达６．０４－６．２４ＬｏｇＰＦＵ／ｍｌ，１代后的各代（至１１代）病毒滴度无明显提高。（３）该细胞接种１４－２株病毒时不产生明显的破坏性病变（ＣＰＥ），在收毒前采取冻溶１次比冻溶前的病毒滴度提高０．２７－０．５ＬｏｇＰＦＵ／ｍｌ。以ＢＤＫ细胞传１代的１４－２株病毒作生产毒种，收毒前冻溶１次等工艺法制备五批疫苗，全面检定结果均符合《乙脑活疫苗规程》的质量标准。其中病毒滴度为６．０９－６．２４ＬｏｇＰＦＵ／ｍｌ；免疫效价（ＩＤ５０＝ｍｌ）为１．９－４．０×１０－５，与相同滴度的原毒株１４－２ＰＨＫ苗对照无明显差异（ｔ＝０．９６８Ｐ＞０．０５），表明其免疫原性与原毒株相似。
Beagle dogs (5 to 10 days old) kidney pieces were digested by heat digestion and the cells were stored in liquid nitrogen. After passing the test, the following tests were performed: (1) After resuscitation and culture, the cells were maintained at 4 Days, overcoming the shortcomings of conventional digestion-cultured cells (36 hours). (2) The virus titer of the first generation virus reached 6.04-6.24LogPFU / ml when passaged 14-2 strains of virus. There was no significant increase in the titer of the virus from the first generation to the eleventh generation. (3) The cells did not produce significant devastating lesions (CPE) when inoculated with 14-2 strains of virus, and the virus titer before freezing was higher than that before freeze-thaw by 0.27-0.5LogPFU / ml . Five batches of vaccine were prepared by the process of producing 14-2 strains of virus of BDK cell passage one generation and frozen-thawing once before harvest. The results of the comprehensive test all met the quality standard of “JE vaccine program”. The virus titer was 6.09-6.24LogPFU / ml; the immune titer (ID50 = ml) was 1.9-4.0 × 10-5, which was the same as that of 14-2PHK seedlings with the same titer Significant difference (t = 0.968P> 0.05), indicating that its immunogenicity similar to the original strain.