Gender-specific neuronal cultures from cryopreserved preparations

All research funded by NIH grants must address a possibility of gender differences in experimental models. This NIH request applies to research conducted on primary neuronal cultures. Since primary neuronal cultures are typically prepared using brain tissue dissected from rodent fetuses, the fetuses need to be genotyped for sex via PCR, the males need to be segregated from the females, and the respective brain tissue needs to be processed separately to obtain specifically male and female brain cells. The genotyping of fetuses for sex greatly extends the time needed to prepare the cultures.

Cryopreserved gender-specific neurons would greatly shorten the time needed to prepare gender-specific primary neuronal cultures. Since no cryopreserved gender-specific neurons are currently available commercially, Spot Cells LLC received a Phase I SBIR grant from the NIH to apply company’s proprietary technology to generate and test gender-specific primary cultures of cortical neurons from mice. Images of cultures at day in vitro (DIV)1-6 from prototype cryopreserved preparations are shown above. The pictures show changes taking place in the same microscopic field overtime.

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