Nutristem xf media

Fibroblasts were sent to either the New York Stem Cell Foundation Research Institute (NYSCF) or Cellular Reprogramming, Inc. (San Diego, CA) for RNA reprogramming. Four SZ and five HC lines were derived at NYSCF and two SZ and one HC lines (converted as part of another study) were from Cellular Reprogramming. NYSCF reprogramming is based on mRNA/miRNA transfection as detailed in Paull et al.^{24 (link)}. Fibroblasts sent to Cellular Reprogramming were similarly converted and the resulting iPS cell colonies were stabilized and expanded. Each fibroblast line was plated to six-well plates without feeders at three different plating densities and subjected to messenger RNA reprogramming. Colonies were bulk-passaged from the most productive well to establish passage 1 iPS cell cultures on rLaminin-521 (BioLamina) in Nutristem XF media (Biological Industries) and expanded in the same culture system until at least passage 3 before being characterized by DAPI/OCT4/TRA-1-60 immunostaining and frozen down. Cells from both sources were made with the same technology and had similar properties.

Human fibroblasts are subjected to mRNA reprogramming [63 (link)–65 (link)] using a cocktail incorporating engineered, chimeric transcription factors to accelerate lineage conversion. The resulting iPSC colonies were stabilized and expanded under xeno-free conditions by Cellular Reprogramming, Inc. (Pasadena, CA). Colonies are bulk-passaged from the most productive well to establish passage 1 iPSC cultures on rLaminin-521 (BioLamina, Sundbyberg, Sweden) in Nutristem XF media (Biological Industries, Kibbutz Beit-Haemek, Israel) and expanded in the same culture system until at least passage 3 before being frozen down for storage. Our iPSC lines are monitored biweekly for mycoplasma infection when in culture and checked for mycoplasma prior to cryopreservation. G-band karyotyping is performed prior to use for downstream experimentation.
The Partners Institutional Review Board (IRB) approved of study activities herein. IRB and FDA IND (investigational new drug) numbers: Harvard Biomarker Study (2003P000541), CLIMB (2017P001169), Fibroblast protocol (2009P000775), Neurobooth (2021P000257), Wrist sensor (2019P002752), experimental PET ligand (2016: 2016P002373, 2020: 2020P003415).