READ ME File for the Article 'A 3D‑induced pluripotent stem cell‑derived human neural culture 
model to study certain molecular and biochemical aspects 
of Alzheimer’s disease.'

Dataset DOI:https://doi.org/10.5258/SOTON/D2630


ReadMe Author: Preeti Prasannan, University of Southampton 

Date of data collection: 2018-2020

Information about geographic location of data collection: UNIVERSITY OF SOUTHAMPTON, UK.


Licenses/restrictions placed on the data, or limitations of reuse:CC-BY


This dataset supports the publication:

AUTHORS:Prasannan, P., Siney, E., Chatterjee, S. et al

TITLE:A 3D-induced pluripotent stem cell-derived human neural culture model to study certain molecular and biochemical aspects of Alzheimer’s disease.

JOURNAL:In Vitro Models

PAPER DOI IF KNOWN:https://doi.org/10.1007/s44164-022-00038-5



This dataset contains: 
1. IMMUNOFLUORESCENCE DATA
2. WESTERNBLOT DATA

1. IMMUNOFLUORESCENCE DATA

This dataset contains all the Immunofluorescence data used to generate the data included in Fig 2 and 3.

Description:

Five iPSC derived NSC cell lines: AD-iPSCs with Presenilin 1 mutation HAD2, HAD3, HAD4 and controls HN8 and HN9 were differentiated into neurons and astrocytes both in 2D and 3D culture systems. The cultures were analysed at different timepoints- day 0, 6weeks (2D and 3D) and 12 weeks (3D) for different markers such as MAP2, Nestin, B3 Tubulin, GFAP, VGLUT 1 and GAD65/67 using Immunofluorescence technique. A Confocal Laser Scanning Microscope-SP8 was used for imaging and ImageJ software was used to measure total positive area of stained cells. Graphpad Prism was used for statistical analysis of the data.


2. WESTERN BLOT DATA 

This dataset contains all the Western Blot data used to generate the data included in Fig 4 from the main text and Online Resource 2 and 3 from supplementary figures. 

Description:

Five iPSC derived NSC cell lines: AD-iPSCs with Presenilin 1 mutation HAD2, HAD3, HAD4 and controls HN8 and HN9 were differentiated into neurons and astrocytes both in 2D and 3D culture systems. Total lysates from these cultures were analysed at different timepoints- 6weeks (2D and 3D) and 12 weeks (3D) for different markers such as Total Tau and PHF1 using Westernblot technique. Fractionated lysates from the 3D cultures at 6 weeks timepoints were analysed to study aggregation of Tau and its isoforms 3R Tau and 4R Tau. Licor Imaging System with Odessey software was used to detect signals. Blots were processed using ImageJ. Graphpad Prism was used for statistical analysis of the data.