In this study, we for the first time carried out Sika Deer (Cervus nippon) somatic cell culture in the space by using self-developed olyvinylidene fluoride (PVDF) membrane culture method, and analyzed the biological characteristic changes of these cells, including cell growth curve and karyotype characteristics. Ear skin tissue was collected for primary adherent culture of somatic cells, and these cells were then subcultured after trypsin digestion. The cells were cryopreserved by using a traditional procedure cooling system. The cell number was counted every day in the 24-well plates and the cell growth curve was made by applying Excel 97-2003. Non-linear regression analysis subroutine Logistic curve model from the IBM SPSS Statistics 23 statistical software was used to analyze growth curve fitting degree, and normal chromosome specimen preparation technology and cytogenetic workstation (AI) software were used to analyze chromosome karyotype of space-treated cells. The results showed that new technology of developed PVDF membrane for cell culture (Fig. 1) could effectively maintain the survival of the C. nippon somatic cells in the space and the cells successfully proliferated after recovery. The somatic cells were still showing normal fibroblast morphology after the space-mutagenesis (Fig. 2). The growth curve of somatic cells showed “S” model which was similar to that of control somatic cells. The space-treated group of cultured somatic cells entered the logarithmic growth phase by 4 days after the cultivation, while it took 2 days in the control group (Fig. 3). The rates of fitting growth curve were similar between two the groups, however, the cell inflexion time and cell volume of somatic cells were significantly changed in space-treated cells (Table 1). Karyotype analysis showed that the chromosome number was still 2n = 66 (XX), and chromosome morphology was normal, 32 of which were autosomes and one pair of sex chromosome (Fig. 4). This study established a new technology of somatic cell culture for domestic animals in the space condition, and also analyzed the effect of the space conditions on biological characteristics of C. nippon somatic cells, which provided some basic and important information and technique for future studies of animal genetics.