Researchers at the Xi’an Children’s Hospital in China have been able to create 2 human-induced pluripotent stem cell (iPSC) lines carrying Pompe disease causing mutations, as reported in Stem Cell Research.

The cell lines were generated from the parents of a child with infantile-onset Pompe disease, from which researchers had also previously created a compound-mutation iPSC line. The previous cell line carried both the GAA gene mutations R608X and E888X together. The 2 new cell lines created from the child’s parents each carry only one of the mutations.

“These 2 lines are valuable resources for studying the pathological mechanisms of GAA mutation-caused Pompe disease,” the study authors said.

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To create the cell lines, peripheral blood mononuclear cells (PBMCs) were collected from the child’s mother and father. The mother carried the GAA mutation of c.2662G > T, p.E888X while the father carried the mutation of c.1822C > T, p.R608X.

The PBMCs were reprogrammed to iPSCs using a Sendai virus containing the 4 Yamanaka factors: OCT4, SOX2, KLF4, and CMYC. The resulting iPSCs exhibited normal morphology as well as normal karyotypes and were found to be mycoplasma-negative.

High expression of the OCT4, SOX2, and NANOG pluripotency markers and the KI67 self-renewal marker were observed through immunostaining. mRNA levels of these markers, as well as the AURKB self-renewal marker, were confirmed through quantitative polymerase chain reactions. Pluripotency was also confirmed through the iPSC’s ability to differentiate into all 3 germ layers.

Real-time PCR confirmed the absence of exogenous reprogramming factors within the cell lines. Sanger sequencing verified that the cell lines contained the heterozygous mutations. Short-tandem repeat analysis demonstrated that the cell lines completely overlapped with those of the donors.

“These two iPSCs lines are an excellent tool for modeling Pompe disease to elucidate underlying pathological mechanisms as heterozygous mutation controls,” the authors concluded.


Huang W, Zhou Y, Wang J, Jiang C, Zhang Y, Zhou R. Generation of two heterozygous GAA mutation-carrying human induced pluripotent stem cell lines (XACHi005-A, XACHi006-A) from parents of an infant with Pompe disease. Stem Cell Res. 2022;64(102934):102934. doi:10.1016/j.scr.2022.102934