Genotyped and Single Cyst-derived Human ADPKD Cell Platforms for Industry and Academia

DiscoveryBioMedIncDBMIncor DBMspecializes in normal and diseased human cell platform engineeringDBM has assembled a consortium of expertswherebycollectivelywe seek Fast Track SBIR funding to establish and offer a Product Line Menu of normal and ADPKD human cell platforms for the academic and industry marketplacesExisting and interested DBM clients will be presented in terms of which options in the Product Line Menu fit their research needsas highlighted in our Research Strategy and Commercialization PlanThis highly anticipated Product Line Menu will include an array ofaspecialty cell culture componentsbprimary normal and ADPKD human primary culturescmixed immortal cultures anddclonal immortal cell lines that are well characterizedgenotypedand defined as to nephron segment of originImmortal cell lines will be compared to the primary cultures from which they were derived to determine whether they are similar in bioassay performancein genotypeand in nephron segment of originDBM and collaborators have begun the collection and characterization ofsingle cyst derived primary cultures andmulticystic tissue derived primary cultures fromhuman ADPKDhuADPKDdonor kidneysprocured from commercial vendorsin pilot Phaselike studiesThese initialhuADPKD primary cultures were combined with primary cultures fromdifferent donors from DrDarren Wallace s PKD Research Biomaterials and Cellular Models Core at Kansas University Medical CenterKUMCso that allsamples could be genotyped by DrPeter HarrisCore at the Mayo PKD CenterEach huADPKD primary culture has been profiled inD microtiter plate driven proliferation bioassays on tissue culture plastic as well as inD Matrigel based cystogenesis bioassaysPreliminary data are presented in support of the above pilot effortsSelect primary cultures that display a compelling genotype and exhibit excellent utility inD andD bioassays will be immortalized by multiple genetic methodsDBM has pledged to also immortalize KUMC primary cultures as necessary in collaborationWhere possiblenephron segment of origin will be identified in huADPKD cellsTwo hypotheses will be addressed within our applied science researchDoes primary huADPKD fibroblast supportco cultureconditioned mediumbothdrive human ADPKD cystic epithelial cell primary cultures to be longer lived and or to attain specific phenotypes inD andD bioassays for PKD researchandDo individual cysts within a given huADPKD donor kidney possess the same genotype or can they differ in genotype from cyst to cysti eexploring theone hitvstwo hithypothesis or the emergence of somatic mutations and the possible presence of haploinsufficiencyDBM et alsetmilestones for our work to be supported by this possible Fast Track SBIR awardMilestonewill be the establishment of single cyst derived and multicystic tissue derived primary cultures from each huADPKD donor kidney tissue sample and with a final goal ofdifferent donors processedMilestonewill be to genotype each individual cyst derived and multicystic tissue derived primary culturecompare genotypes against the ADPKD genotype databaseand select candidate primary cultures for immortalizationMilestonewill be to immortalize select primary cultures using a well optimized Critical PathMilestonewill be to characterize immortal clonal cell lines versus primary cultures wherefrom the cell lines originated to determineusabilityin different bioassaysMilestonewill be to offer custom packages of human normal and ADPKD kidney cell platforms from our Product Line Menu to the PKD academic and industry research communities in Yearsandof the award and beyond this term as a self sustaining business