Here, we isolated two novel rpESCs on 28% efficiency, comparable to biparental counterpart. These rpESCs shared many properties with other primate ESCs, including expression of pluripotent markers, capacity to generate derivatives representative of all three germ layers in vivo and in vitro, maintaining of euploid karyotype even after long culture. Because of lack of paternal genome, paternally expressed imprinted genes were expected to be absence in rpESCs. However, in our two novel rpESCs, paternally expressed imprinted genes including PEG3, SGCE, PEG10, ZIM2 and MEST could be detectable at least in one rpESCs, consist with other available parthenogenetic rhesus monkey ESCs 
. Nevertheless, SNPRN, NDN and MAGEL2 were absent from both our rpESCs and others 
. Although twelve SNP loci we analyzed were uniform between rpESCs and their oocyte donors, Pa2.2 represented heterozygousity to some extent. This phenomenon recently observed also in mouse and human parthenogenetic ESCs 
, due to homologous crossing-over and recombination during meiosis I.
In our two rpESCs, mir-302 cluster had a very high expression level, consisting with previous studies that mir-302 cluster was high-riched in human ESCs 
. Intriguingly, mir-302 cluster can reprogram cancer or somatic cells into iPS 
revealed that mir-302 cluster had important roles in maintaining pluripotency of pluripotent cells. Recently, the roles of mir-302 cluster in regulation of ESCs pluripotency have been clarified on some extent. Nanog, Oct3/4, Sox2 and Rex1 are upstream regulators of the miR-302 cluster promoter 
, miR-302a regulate ESCs cell cycle by repressing cyclin D1 
, and miR-302 impair early differentiation by repress NR2F2 
and Lefty 
at the post-transcriptional level. On the other hand, miR-302 can enhance reprogramming by accelerating mesenchymal-to-epithelial translation (MET) 
and suppressing some epigenetic regulators 
. MiR-106a-363 cluster and miR-106b-25 cluster, which were also high-riched in rpESCs, were reported to enhance reprogramming by accelerating MET 
. Interestingly, miR-371–373 cluster and C19MC miRNAs, which were reported to be highly enriched in human ESCs 
, only represented a moderate or almost undetectable expression level in our rpESCs respectively. Additionally, some members of let-7 family were upregulated in rpESCs. These results were also observed in some human ESCs lines 
. This variance in some miRNAs may reflect difference in culture condition, developmental stage when ICM were isolated.
Although rpESCs shared many properties with their IVF counterpart, comparing between them on miRNA level would help understanding universality and particularity of rpESCs. For miRNAs which had a very high expression level(>10000 reads TPM) in both parthenogenetic and IVF rhesus monkey ESCs, members of mir-302 cluster, miR-106b-25 cluster have important roles in maintaining of pluripotency. Interestingly, Guo and colleagues recently reported 
that members of miR-99b-125a cluster increased the number of hematopoietic stem cells in vivo by inhibiting pro-apoptotic genes Bak1. This cluster was also up-regulated expression in our rhesus monkey ESCs despite of origin from parthenogenesis or IVF, suggesting the potential roles of this cluster in ESCs and IPS. Consisting with Laurent and colleagues' reports 
, miR-183–182 cluster, miR-103, miR-21 and miR-378, which were reported to be related to many type carcinomas 
, were also enriched in both types of rhesus monkey ESCs. Our results revealed that rpESCs and IVF-origin rhesus monkey ESCs represented high similarity with respect to comprehensive miRNA expression pattern, especially for miRNAs which involved in properties of stem cells. Since parthenogenetic ESCs were considered as potential source for regeneration medicine, their tumorigenesis risk should be comprehensively and carefully evaluated. By comparing with IVF-origin rhesus monkey ESCs, our results indicated that rpESCs would up- or down-regulated some specific tumor-related miRNAs, in addition to up-regulating of some oncogenic miRNAs in both type of rhesus monkey ESCs mentioned above. Oncogenic miRNAs 
, including let-7 family, miR-374 family, miR-138, miR-195, miR-19a and miR-200a, had more than twice expression level in rpESCs comparing with their IVF counterparts. Reversely, tumor suppressor, including miR-222, miR-342-3p, were remarkably down-regulated in rpESCs. These results, along with recent report that cell lines from human parthenogenetic embryos had aberrant centriole distribution 
, highlighted the tumorigenesis risk of parthenogenetic ESCs.
X-chromosome inactivation, by which dosage compensation of the sex chromosome is achieved, is widely presented in mammals 
. In mouse embryonic stem cells (mESCs) and mouse-induced pluripotent stem cells (miPSCs) both X chromosomes are active, and inactivation occurs during differentiation 
. The presence of two active X chromosomes have been considered as one of epigenetic markers for murine ESCs. However, in human ESCs or iPS, X chromosomes status represented complex variations 
. Especially, by changing culture and growth factor condition, pre-X inactivation human ESCs or iPS could be derived 
. By collecting and analysing the expression of the entire set of genes on the X chromosome, Bruck and colleagues divided human pluripotent cell lines into three categories: no XCI, full XCI, and partial XCI 
. In current study, we collected and analyzed the expression of the entire set of known miRNAs on the X chromosome in our rhesus monkey ESCs. The X-linked miRNAs in Pa3 rpESCs had about twice level of expression than that of Pa2.2 rpESCs and their IVF counterparts, implying the activation of both two of X chromosome in Pa3. In order to validate this prediction, we checked XIST, a non-code RNA which was specifically expressed in female cells and took part in XCI. Expectably, XIST could not be detected in Pa3 rpESCs and male rhesus monkey ESCs IVF3.3. Whereas, Pa2.2 expressed high level of XIST, equal to female rhesus monkey ESCs IVF3.2. These results suggested that the variations of X chromosome status were also presented in parthenogenetic ESCs, regardless of the identical origin of both X chromosome. Notably, in Rhesus monkey, both X chromosomes are active in blastocyst stage from which ESCs were isolated 
. Therefore, XCI would occur during derivation of ESCs.