Data on the effect of oral feeding of Arachidonic acid or Docosahexanoic acid on haematopoiesis in mice

Stem cells have peculiar property to self-renew and differentiate. It is important to control their fate in safe and effective ways for their therapeutic use. The mediators of essential polyunsaturated fatty acids (PUFAs) namely Arachidonic acid (AA) and Docosahexanoic acid (DHA) are known to play a role in haematopoiesis via various metabolic pathways [1]. However the direct effect of purified AA or DHA on haematopoiesis has not been well investigated yet. We have reported that oral administration of PUFAs enhanced haematopoiesis in mice [2]. Signaling Leukocyte Antigen Molecule (SLAM) (CD48−CD150+) phenotype consists of pure population of haematopoietic stem cells (HSCs). Herein we observed higher percentage of SLAM (CD48−CD150+) phenotype in the bone marrow (BM) cells of mice fed with AA or DHA compared to PBS fed control mice. Data from engraftment study depicts that BM from AA/DHA-fed mice showed higher absolute number of donor cells in recipient mice compared to control. The enhanced hematopoiesis observed in AA/DHA-fed mice was returned to normal when the mice were kept on normal diet for six weeks (after ten days of oral feeding). We confirmed GCMS (Gas Chromatography-Mass Spectroscopy) retention times of AA and DHA by co-injecting fatty acid extract from AA or DHA fed mice with purified AA or DHA standards respectively. Representative flow cytometry profile of Lin−Sca-1+c-kit+(LSK) cells showed higher expression of CXCR4 protein and ligands of Wnt, Notch1 signaling in BM of AA/DHA-fed mice.


a b s t r a c t
Stem cells have peculiar property to self-renew and differentiate. It is important to control their fate in safe and effective ways for their therapeutic use. The mediators of essential polyunsaturated fatty acids (PUFAs) namely Arachidonic acid (AA) and Docosahexanoic acid (DHA) are known to play a role in haematopoiesis via various metabolic pathways [1]. However the direct effect of purified AA or DHA on haematopoiesis has not been well investigated yet. We have reported that oral administration of PUFAs enhanced haematopoiesis in mice [2]. Signaling Leukocyte Antigen Molecule (SLAM) (CD48 − CD150 þ ) phenotype consists of pure population of haematopoietic stem cells (HSCs). Herein we observed higher percentage of SLAM (CD48 − CD150 þ ) phenotype in the bone marrow (BM) cells of mice fed with AA or DHA compared to PBS fed control mice. Data from engraftment study depicts that BM from AA/DHA-fed mice showed higher absolute number of donor cells in recipient mice compared to control. The enhanced hematopoiesis observed in AA/DHA-fed mice was returned to normal when the mice were kept on normal diet for six weeks (after ten days of oral feeding). We confirmed GCMS (Gas Chromatography-Mass Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/dib Spectroscopy) retention times of AA and DHA by co-injecting fatty acid extract from AA or DHA fed mice with purified AA or DHA standards respectively. Representative flow cytometry profile of Lin − Sca-1 þ c-kit þ (LSK) cells showed higher expression of CXCR4 protein and ligands of Wnt, Notch1 signaling in BM of AA/DHA-fed mice.
& 2017 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Subject area
Biology More specific subject area

Nutritional metabolism, Nutritional Biochemistry
Type of data Graph, figure How data was acquired Flow cytometry, Gas chromatography-mass spectroscopy (GC-MS)

Data format
Analyzed data Experimental factors C57/BL6 mice were fed with AA/DHA daily for ten days and their bone marrow cells were subjected to various assays for haematopoiesis Experimental features Bone marrow cells of mice were stained with specific antibodies for HSCs were analyzed by immunofluorescence method, For engraftment study, bone marrow cells from AA/DHA-fed mice were infused in irradiated congenic recipient mice and donor population was analyzed.

Data source location
Stem cell laboratory, Lab #4, National Center for Cell Science, Savitribai Phule Pune University Campus, Pune, India Data accessibility Data is available with this article

Value of the data
This data set is of value to the researchers seeking effect of oral administration of AA or DHA on haematopoiesis (phenotypic as well as in vivo engraftment).
Data could facilitate analysis of GCMS (Gas Chromatography-Mass Spectroscopy) chromatograms of AA/DHA. Data shows that Wnt, Notch1 and CXCR4 signaling are probable molecular mechanisms being activated in enhanced haematopoiesis of AA/DHA-fed mice.

Data
The data give comparative account of bone marrow (BM) cells of mice fed with AA/DHA with control mice fed with saturated fatty acid (SFA) -Palmitic Acid. Data of phenotypic flow cytometry analyses as well as statistics of bone marrow cells of mice fed with AA or DHA with control mice, is shown. The data also give graphical representation of various parameters for haematopoiesis of mice kept on normal diet after ten days feeding of AA or DHA( Fig.1 and Fig.2). Comparison of absolute numbers of engrafted donor cells from bone marrow of AA/DHA-fed mice is represented in the statistical format (Fig. 3).Comparative account between mice fed for ten days & sacrificed on next day and mice fed for ten days and sacrificed after six week, is shown (Fig. 4). The data of co-injection of fatty acid samples from bone marrow from AA or DHA fed mice with that of pure standards of AA and DHA is shown (Fig. 5). Data give representative flow cytometry profiles of bone marrow of AA/DHAfed mice for ligands of Wnt and Notch1 signaling and CXCR4 protein (Fig. 6).

Oral feeding of mice
C57/BL6 (6-8 weeks) mice were fed daily through oral feeding gavage for ten days with 8 mg of either AA or DHA/mouse/day. PBS fed mice were kept as control. Mice fed with 8 mg of Palmitic acid were kept as additional control in initial experiments. Their BM cells were harvested and were subjected to assays for haematopoiesis. Detailed experimental design is described in Limbkar et al. [2].

SLAM marker expression analysis
LSK analysis was performed as per Limbkar et al. [2]. For SLAM (CD48 − CD150 þ ) antigen detection, LSK cells of PBS/AA/DHA fed mice were further stained with CD48APC-Cy7 and CD150 BV421. Cells were analyzed by flow cytometry.

Engraftment studies
This assay was done as described by Spangrude and coworkers, 1995. The detailed methodology is described in Limbkar et al. [2].
Absolute numbers of engrafted donor cells in recipient mice were calculated by the formula below.

Co-injection experiment
Methyl esters of AA and DHA standards were co-injected with methyl esters from BM cells of AA and DHA fed mice and analyzed by GC-MS. Methyl esters were prepared as per Limbkar et al. [2]

Transparency document. Supporting information
Transparency data associated with this article can be found in the online version at http://dx.doi. org/10.1016/j.dib.2017.08.009.